www.csiro.au
15th Annual Protein Expression Workshop
Proteins for structural
and biological studies
30 July – 1 August 2014
https://wiki.csiro.au/display/PEWS/PEWS+Home
Major
sponsors
www.csiro.au
15th Annual Protein Expression Workshop
Proteins for structural and biological studies
Introduction
The CSIRO Protein Expression Workshop is an annual event
designed to improve the understanding of proteins for structural
and biological studies. Join us for the 15th Annual Protein
Expression Workshop from 30 July – 1 August 2014.
Workshop aims
The production of complex proteins for structural and biological
studies requires a range of skills and disciplines including
molecular biology, fermentation, protein purification and
structural analysis. This workshop will cover a range of topics
to enable successful production work and provides an excellent
networking opportunity.
By the end of the Workshop
participants will:
• have an understanding of different expression
systems
• be familiar with basic technologies involved in
protein production
• know the most important large and smallscale equipments involved in expression and
purification
• be able to design and optimise your production
system
• understand basic purification and assay methods
• learn how to integrate your production system
with purification
• be familiar with current trends in protein
production and fermentation
https://wiki.csiro.au/display/PEWS/PEWS+Home
• learn how collaborative facilities might be able to
assist you to reach your research goals.
Major sponsors
Gold sponsors
Silver sponsors
b
Blueprints for a sustainable future
Bronze sponsors
www.biosci.com.au
Hands-on Workshop sponsored by
4
Kinesis
Biorad
POCD
Scientific
Invitro
food
TRENDBIO
VWR
Lecture
theatre
ATA
Scientific
ThermoFisher
SARTORIUS
EPPENDORF
Life
Life
technologies technologies
GE
healthcare
BIONOVUS
Bioscientific
MERCK
Millipore
Millennium
Science
Corridor
LabTek
d
o
o
f
e
e
f
f
o
c
&
a
e
t
2nd floor at Parkville, C S I R O
PALL
3M
Purification
3M
Purification
TRENDBIO
iScience
LONZA
Grace
Sponsor’s area for the 15th Protein Expression Workshop, 30/07 – 01/08/2014
GENSEARCH
windows
15th Annual Protein Expression Workshop
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Program
DAY ONE : Wednesday 30th July 2014
Venue
CSIRO - 343 Royal Parade, Parkville
8:30AM
Registration
9:15AM
Safety induction and housekeeping
SETTING THE SCENE
Chair: Greg Coia
9:20AM
Introduction and welcome
Paul Savage
(Manufacturing Flagship)
9:30AM
OPT-302: A novel VEGFR-3 fusion protein for the
treatment of wet AMD
Mike Gerometta
(Circadian Technologies Ltd)
10:00AM
Non-native and modified particulate immunogens
with enhanced immunogenicity
Hans Netter (Monash University)
10:30AM
MORNING TEA
11:00AM
Helicobacter: Then and Now
Barry Marshall
(University of Western Australia)
12:00PM
Rotavirus vaccine development
Rajendra Sabale
(Serum Institute of India)
12:30PM
LUNCH
PROTEIN EXPRESSION
Chair: Ian Macreadie
1:30PM
Systems for large-scale manufacture of vaccines and
biopharmaceuticals
John Power (Zoetis)
2:00PM
Strategies for expressing recombinant proteins in
bacteria and their purification
Bill McKinstry (CSIRO)
2:30PM
Improving Secretion and Purification of
Glycoproteins in the Baculovirus-Insect Cell
Expression System
Cindy Chang (UQ-PEF)
3:00PM
Yeast expression systems
Brendon Monahan (CSIRO)
3:30PM
AFTERNOON TEA
4:00PM
Mammalian toolbox – Platform technology for
developing human therapeutics
Jeff Hou (AIBN)
4:30PM
Discovery and development of novel human
therapeutics using recombinant protein expression
in mammalian cells
CareerLounge: Connecting students, employers and
educators
Catherine Owczarek (CSL)
5:00PM
5:30PM
Dominique Fisher (CareerLounge)
Discussion and close of Day 1
DRINKS AND PIZZAS
Sponsored by BPN
15th Protein Expression Workshop • https://wiki.csiro.au/display/PEWS/PEWS+Home
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15th Annual Protein Expression Workshop
Program
DAY TWO : Thursday 31st July 2014
Venue
CSIRO - 343 Royal Parade, Parkville
9:00AM
Registration
9:25AM
Welcome
Tim Adams (CSIRO)
9:30AM
Control of Glycosylation Profiles in Cell Bioprocesses
Mike Butler (Univ of Manitoba)
PROTEIN PURIFICATION & CHARACTERISATION Chair: Lou Fabri
10:00AM
Purification and characterisation of mammalian expressed
proteins: key assessment considerations for
candidate/target selection in a research based
commercial environment
10:30AM
MORNING TEA
11:00AM
Structural Biology at the Australian Synchrotron
Jose Varghese (CSIRO)
11:30AM
Simple and low cost quality control of expressed
proteins by mass spectrometry
Nick Williamson
(Bio21/Melbourne University)
12:00PM
Physical techniques in a protein workflow
Shane Seabrook (CSIRO)
12:30PM
LUNCH
Pierre Scotney and Peter
Schmidt (CSL)
COLLABORATIVE CENTRES & SUCCESS STORIES
Chair: Jan Tennent
2:00PM
Australia Therapeutics Pipeline
Stewart Hay (TIA)
2:15PM
The National Biologics Facilities (NBF) – A joint
AIBN/CSIRO Facility
Peter Gray (AIBN/UQ)
2:30PM
BPA and the Biologics Facility of the Future
Brett Whitecross (BPA)
2:45PM
The Victorian Antibody Initiative (VAI)
Caroline Laverty (MATF)
3:00PM
GMP Considerations for recombinant protein
manufacture and working with a CMO
Brett Butcher (PharmaSynth)
3:30PM
AFTERNOON TEA
4:00PM
Insulin receptor: production of midi-, mini- and
micro-constructs: Targeted design of insulin receptor
for crystallography of Site 1
John Menting (WEHI)
4:30PM
Telomerase: a promising target for cancer
Scott Cohen (CMRI)
5:00PM
Discussion and close of Day 2
5:30PM
PRE-DINNER DRINKS
7:00PM
CONFERENCE DINNER
Fish Bowl at the Melbourne Aquarium
Sponsored by
BioMedVic
15th Protein Expression Workshop • https://wiki.csiro.au/display/PEWS/PEWS+Home
6
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Program
DAY THREE : Friday 1st August 2014
Venues
CSIRO Ian Wark Lab Reception
Bayview Avenue, Clayton
CSIRO Parkville Laboratories
343 Royal Parade, Parkville
8:40AM
Meeting at Parkville reception for
bus ride to Clayton workshops
n/a
9:00AM
n/a
Welcome to Parkville & Safety Induction
(2nd floor Seminar room)
9:15AM
Welcome to Clayton & Safety
Induction (Ian Wark Canteen)
WORKSHOP STARTS
9:30PM
10:40AM
MORNING TEA
11:00PM
WORKSHOP CONTINUES
12::30PM
LUNCH
1:30PM
WORKSHOP CONTINUES
3:00PM
AFTERNOON TEA
3:15PM
WORKSHOP CONTINUES
4:00PM
Bus leaves to go back to Parkville
4:30PM
CLOSE OF WORKSHOP
WORKSHOP 1: Microbial Scale-up
Bioreactor assembly, probe calibration,
sampling and process monitoring. Including
demonstration of the Sartorius BioStat B
benchtop bioreactor.
Geoff Dumsday
Linda Howell
Andreas Kocourek
Kanika Bhadoria
WORKSHOP 2: Mammalian Cell Culture
Mylinh La and Louis Lu
ReadyToProcess WAVE™ 25 (GE)
John Ince
Cytell Cell Imaging System (GE)
Tracee Archibald
Chemidoc for V3 Western Workflow (BioRad)
Bansi Sanghvi
Transient transfection/Cell count and analysis
(Life Technologies)
Alison Digney and Naomi Morisson
Single-use Bioreactor system (Eppendorf)
Stephen de Lacey
n/a
CLOSE OF WORKSHOP
WORKSHOP 3: Protein Purification and
Characterisation
Participants will have the chance to attend 2 of
the following 6 workshops:
Mini Workshop 1:
Akta-Xpress (GE)
Lesley Pearce/David Cossens
(3h)
Mini Workshop 2:
Profinia/Duo-Flow (BioRad)
John Bentley/Nishen Naidoo
(3h)
Mini Workshop 3:
His-tagged protein purification
Pat Pilling
(3h)
Mini Workshop 4:
Mass Spectrometry
Lindsay Sparrow/Nick Bartone
(1.5h)
Mini Workshop 5:
Size-Exclusion Chromatography (ATA)
Paul Barrett
(3h)
Mini Workshop 6:
BLItz Sytems (Millenium Science)
Mike Yarski
(3h)
15th Protein Expression Workshop • https://wiki.csiro.au/display/PEWS/PEWS+Home
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14th Annual Protein Expression Workshop
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PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day One
Presenters
Introduction and welcome
Notes
PAUL SAVAGE
CSIRO Manufacturing Flagship
Dr Paul Savage is the research director for the Biomanufacturing Program in the
Manufacturing Flagship at CSIRO. His program provides scientific capabilities in support
of companies involved in biotechnology and chemical innovation. This includes the
design, construction and process tech transfer for biological and synthetic molecules of
potential commercial importance.
The program primarily uses the science capabilities of:
•molecular and cell biology including enzymology
•stem cell biology and scale-up
•bacterial and mammalian cell fermentation up 500L scale and downstream
processing
•protein science including structural biology and X-ray crystallography
•synthetic organic chemistry including scale up and process chemistry
•flow chemistry and high-throughput synthesis
Paul is working at the interface between business and science to help build a vibrant,
globally successful Australian biotech industry.
OPT-302: A novel VEGFR-3 fusion protein for the
treatment of wet AMD
Mike Gerometta
Circadian Technologies Ltd
OPT-302 is a soluble form of the vascular endothelial growth factor receptor-3 (VEGFR-3)
expressed as an Fc-fusion protein. OPT-302 functions by capturing and thereby
neutralizing the activity of VEGF-C and VEGF-D on the endogenous receptors VEGFR-2
and VEGFR-3. OPT-302 is highly specific for VEGF-C and VEGF-D, and does not bind
VEGF-A. Administration of OPT-302 can inhibit blood and lymphatic vessel growth,
as well as vessel leakage, which are characteristic hallmarks of several eye diseases,
including neovascular (“wet”) macular degeneration (AMD).
The molecular targets for OPT-302, VEGF-C and VEGF-D, differ from that of marketed
biologic VEGF-A inhibitors, such as ranibizumab and aflibercept, which bind VEGF-A
and prevent the interaction of VEGF-A with its receptors (VEGFR-1 and VEGFR-2) on the
surface of endothelial cells, reducing endothelial cell proliferation, vascular leakage,
and new blood vessel formation. Therefore, the therapeutic activity of OPT-302 may be
complementary to that of existing therapies for eye disease.
OPT-302 has been designed as an optimal ophthalmic agent for intravitreal
administration. It has high affinity for its cognate ligands, a long ocular half-life
potentially enabling a reduced administration regime, rapid blood clearance to
minimize systemic adverse events and employs endogeneous proteins for reduced
immunogenicity. The molecular structure also facilitates protein expression and
purification. These properties have been displayed in pre-clinical models which show
that OPT-302 inhibits wet AMD either alone or in combination with existing drugs.
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15th Annual Protein Expression Workshop
Day One
Notes
Biography
Dr Mike Gerometta has been Head of Chemistry, Manufacturing & Controls (CMC),
Development since 2008 with responsibilities encompassing the outsourcing of
Circadian’s research and cGMP manufacturing activities for its therapeutic and diagnostic
portfolio, as well as oversight of internal laboratory activities. Mike over 25 years’
experience in the Australian biotechnology industry, most recently as Chief Operating
Officer of Q-Gen, the manufacturing facility of the Queensland Institute of Medical
Research. He has extensive experience working with numerous Contract Manufacturing
Organisations overseas and locally in all facets of translational CMC from concept
through to Phase II studies, in the process successfully guiding the manufacture of four
biologics through to Phase 1/2 clinical trials, including oversight of two non-clinical
programs, as well as associated regulatory interactions in North America and Australia.
He has also directed the development of numerous in vitro diagnostic products through
to the market over 19 years at Agen Biomedical, ultimately as Research and Product
Development Director.
Mike was awarded his PhD in biotechnology from the Queensland University of
Technology and has a degree in chemistry from the University of Technology in Sydney.
Non-native and modified particulate immunogens with
targeting capability and enhanced immunogenicity
Hans J. Netter
Monash University
Virus-like particles (VLPs) are tools of a leading innovative bio-nanotechnology in vector
and vaccine development. VLPs composed of the small envelope protein (HBsAgS)
from hepatitis B virus represent a class of bio-nanoparticles, which are formed by viral
structural proteins providing a high density display of antigenic sequences. The capacity
to serve as carriers of antigenic sequences derived from either the parental virus or
foreign sources has broadened their potential as prophylactic and therapeutic vaccine
candidates. They have the ability to trigger key pathways of the immune system accessing
major histocompatibility class I and II specific antigen presentation in dendritic cells.
Novel strategies to manipulate or to enhance immunogenicity by utilizing biochemically
modified VLPs have been developed by changing the level of disulfide bonding and by
modifying the level of glycosylation. VLPs composed of biochemically modified HBsAgS
subunits are utilized to illustrate a possible general principle that extends beyond
hepatitis B research and encompasses strategic developments to manipulate immune
responses by using non-native immunogens to promote significant benefits to human
health.
To allow cell- and tissue-specific approaches, VLPs with targeting ability were designed
and generated. The recombinant VLPs are decorated with functional antibody domains
that specifically target immune cells providing superior immunogenicity compared to
the native (wildtype) VLPs. The presence of functional antigen-binding domains arrayed
on VLPs will allow the development of universal molecular devices with specificities
determined by the selected antibody domains.
Biography
Hans Netter completed his doctorate at the University of Munich, and Max-Planck
Institute for Biochemistry, Germany in 1990 in the field of “autoimmunity”. In the
following year, he joined Prof. John Taylor’s laboratory at Fox Chase Cancer Center,
Philadelphia, USA to investigate the genome stability and helper independent infections
of the hepatitis delta satellite. Research positions at the Medical School, University of
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PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day One
Hamburg, Germany and Medical Virus Research Centre, Brisbane, Australia followed.
He discovered novel avian hepatitis B viruses and non-conventional human hepatitis
B viruses, which demonstrated a functional and evolutionary diversity of hepatitis B
viruses. Due to these achievements, he was awarded a higher doctorate by the Medical
School, University of Hamburg in 1997. In 2002, Hans Netter joined Monash University
as Principal Investigator and Lecturer, with a research focus on immunogen - immune
system interactions. He was awarded the Graduate Certificate in Higher Education in
2004. He has responsibilities in under- and post-graduate teaching and acts as the
Deputy Assoc Dean (Research Degrees), Faculty of Medicine Nursing and Health Sciences,
Monash University.
Notes
Helicobacter: Then and now
Barry Marshall
University of Western Australia, Nobel laureate
The early work on Helicobacter began with simple questions. “Were these bacteria living
in the stomach?” and secondly, “How could they survive in the presence of stomach
acid?” The bacteria were associated with gastritis, so tightly that the cause and effect
hypothesis could not be understated. For a century, some experts had proposed that
gastritis could lead to duodenal and gastric ulcer. The epidemiology also suggested
that gastric cancer was also part of the picture. If gastritis could be controlled then
stomach cancer could be prevented. Various paradoxes existed which were explained
by the natural history of Helicobacter infection, which was unravelled by a volunteer
experiment published in the April 1985 edition of the Medical Journal of Australia.
If the bacteria colonised most of the human race, how long had they been there? Were
they recent, or had they been in humans for millennia? If they had been infecting almost
all humans since the stone-age, did they actually have a useful purpose?
New data have revealed that Helicobacter can have a benefit in several ways, perhaps
assisting the immune system to achieve the correct balance. Some of the most common
human diseases affecting modern humans are due to a hyper-reactive immune system
causing troublesome diseases such as asthma and allergic eczema. In some studies, these
conditions are less common when a Helicobacter infection is present.
Out of curiosity, new genomic technologies have been applied to the Helicobacter
genome. Amazingly, every human race carries a unique strain of Helicobacter and we can
trace its origins back to determine our human origins.
Biography
In 2005 Barry J. Marshall and J. Robin Warren were awarded the Nobel Prize for
Physiology or Medicine in recognition of their 1982 discovery that a bacterium,
Helicobacter pylori, which affects 50% of the global population, causes one of the most
common diseases of mankind, peptic ulcer disease.
Barry Marshall met Robin Warren, a pathologist interested in gastritis, during internal
medicine fellowship training at Royal Perth Hospital in 1981. The pair studied the
presence of spiral bacteria in association with gastritis. The following year (1982),
Helicobacter pylori was cultured for the first time and they developed their hypothesis
related to the bacterial cause of peptic ulcer and gastric cancer.
In 1984, at Fremantle Hospital, Marshall proved that the germ was harmful in a wellpublicised self-administered experiment, in which he drank a culture of H. pylori.
Persevering despite widespread skepticism, Marshall also came up with combinations of
drugs that killed the bacteria and eliminated ulcers permanently.
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15th Annual Protein Expression Workshop
Day One
Notes
In 1984 the World Health Organisation recognized H. pylori as the main cause of stomach
cancer. Marshall and Warren’s work is acknowledged as the most significant discovery in
the history of gastroenterology and is compared to the development of the polio vaccine
and the eradication of smallpox.
In 1998 Marshall was made a Fellow of the Royal Society. In 2008 he was elected as a
Foreign Member of the prestigious US National Academy of Science, an institution that
was established in 1863 by President Abraham Lincoln.
Dr Marshall founded Ondek Pty Ltd in 2005, to develop and commercialise a novel drug
and vaccine delivery platform, based on the use of genetically modified Helicobacter
pylori. Dr Marshall and his research team have developed a set of proprietary
technologies, referred to as the Helicobacter pylori Platform Technology (HPPT). The
HPPT has potential application for oral delivery of immunotherapies, biopharmaceuticals
and other drugs, and vaccines. The first product being developed at Ondek is an
immunotherapy for the treatment of allergic asthma and food allergies in children.
Barry Marshall was born in Kalgoorlie in 1951 and attended Marist Brothers College in
Perth from 1960-68. He completed his undergraduate medical degree at The University
of Western Australia in 1974. He is married with four children and five grandchildren and
lives in Shenton Park, Western Australia.
Rotavirus vaccine development
SABALE RAJENDRA NARAYAN
Serum Institute of India
Serum Institute of India Ltd. (SIIL) is a private organization of Dr. Cyrus Poonawalla
group. SIIL is world’s largest producer of vaccines in terms of number of doses. It is
estimated that two out of every three children from about 140 countries are vaccinated
by SIIL’s vaccine.
At present, Rotarix® and RotaTeq® are the only commercially available Rotavirus
vaccines. Rotavirus vaccine is found promising and cost effective solution to control the
rotavirus diarrhea. The present vaccines are stored and supplied at cold conditions of
2-8°C. SIIL has developed a vaccine with objective that the vaccine should be stored and
transported without cold chain.
SIIL Rotavirus vaccine is live, oral, lyophilized pentavalent vaccine, comprises of five
serotypes viz. G1, G2, G3, G4 and G9 and individually grown on Vero cells. All these
re-assortant strains have VP7 gene of respective serotype of human strains and other
ten genes of bovine (UK) Rotavirus strain. The vaccine is supplied along with 2.5 ml of
diluent (citrate bicarbonate buffer) for reconstitution of lyophilized vaccine.
The stability of vaccine was studied at 2-8°C and 25°C for 36 months, 37°C for 24 months
and 40°C for 6 months. The data suggested that the vaccine was stable for 36 months
at 2°C to 8°C and 25°C. At 37°C and 40°C, the SIIL Rotavirus vaccine was stable for 24
months and six months respectively. This indicates that SIIL’s rotavirus vaccine is thermostable preparation, which may be used at ambient conditions.
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PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day One
Biography
Notes
In my current role as Deputy Manager (Serum Institute of India Ltd, Pune) I am involved in
the planning and execution of the development of rotavirus vaccine. The vaccine is being
developed for the pediatric use to prevent severe diarrhoea in infants. Vaccine Development
work involves the Mammalian cell culture work, virus characterization and formulation
development. The associated activities involve clinical trials and clinical research.
My past research interests included the development of Pod boarer resistant variety of
Chickpea (Cicer arietium, L), via gene transformation, at the Plant Tissue Culture Division
of the National Chemical Laboratory. At Mahyco Life Sciences Research Centre, I had
worked to develop a pest-resistant cotton variety.
I completed both my undergraduate B.Sc. degree in the 1995 and my M.Sc. in 1997 at the
University of Pune, India.
Systems for large scale manufacture of vaccines and
biopharmaceuticals
john power
Zoetis
This presentation will provide an industrial perspective on the manufacturing platforms
for commercial production of vaccines and biopharmaceuticals. While the majority of
biopharmaceuticals approved for human use are monoclonal antibodies produced in
mammalian expression systems, there is no “One-Size-Fits-All” solution for manufacture
of biological medicines which then drives a degree of diversity in manufacturing
platforms across the industry. This diversity is shaped by the inherent technical
requirements of the producer-organism, the desire for improved product safety profiles,
and the ever-increasing demand for cost-efficient production.
A survey of manufacturing platforms will be presented along with commentary on the
technical issues encountered in their industrialisation.
Biography
John Power is a Director of Development in the Veterinary Medicine Research and
Development division of Zoetis. In this role, John provides leadership towards the
industrialisation of new veterinary vaccines in both the Asian and Global pipelines for
new product flow. John has over 20 years post-doctoral experience in the scale-up of
manufacturing systems for commercial production of vaccines and biopharmaceuticals.
He has particular expertise in scale-up of microbial fermentations, cell culture systems,
and associated processing for manufacture of biologicals. He holds a bachelor’s degree
in Applied Science, a Masters in Biotechnology, and a PhD in Chemical Engineering
awarded by the University of Queensland. He has also completed an MBA in Technology
Management. John is a member of ISPE and AusBiotech. Before joining Zoetis, John held
senior research positions in CSL Ltd, Bioproperties (Australia) P/L, Burns Philp, and Fort
Dodge.
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15th Annual Protein Expression Workshop
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Notes
Strategies for expressing recombinant proteins in bacteria
and their purification
bill mckinstry
CSIRO Manufacturing Flagship
So you want to produce a recombinant protein in bacteria? Where to start - what type of
protein is it and is the biological function of your protein known? What do you require
the recombinant protein for? Literature and bioinformatics reviews and website tools.
Designing your protein expression construct, including the use of tags/fusion proteins
to facilitate protein folding and purification. Selecting a suitable host cell to express
your recombinant protein, and what the different protein expression strains do. Using
a medium throughput approach we can rapidly screen protein expression, protein
extraction and protein purification strategies for increasing the production of soluble
recombinant proteins. By exploring different bacterial strains, types of culture media
and induction conditions we are able to increase the likelihood of producing soluble
proteins. What to do if your protein is insoluble. Scaling-up protein expression, and
tips for optimising protein purification and down-stream processing with the view to
delivering quality recombinant proteins.
Biography
Bill McKinstry is a protein biochemist with over 25 years technical expertise in the
expression, purification and analytical characterisation of recombinant proteins
for structural and functional studies. He has experience in academia and both the
biotechnology and pharmaceutical industries. His research interests include; cytokines
and their receptors; membrane associating proteins, host-pathogen interactions, viral
structural proteins – self assembly and macromolecular complex formation, structural
genomics and high-throughput technologies. He is the author of 38 peer-reviewed
publications (including publications in Cell and Nature Structural Biology), has >1700
citations with an H-factor of 20, the inventor of 3 patents of which two have been
licensed to pharmaceutical companies, and the development of downstream processing
protocol for the production of a biotherapeutic recombinant protein about to
commence human clinical trials. He holds Honorary Research Fellow appointments with
both The University of Melbourne and Deakin University.
Improving secretion and purification of glycoproteins in the
baculovirus-insect cell expression system
Cindy Chang
The University of Queensland, Protein Expression Facility
Secretory glycoproteins are required for diverse research applications including
therapeutic and vaccine development. The baculovirus-insect cell expression system is
emerging as a favourable system for the production of recombinant glycoproteins due to
its eukaryotic protein processing capabilities. Low secretion efficiency and downstream
recovery are often the bottlenecks for this system. To improve protein secretion, varying
expression conditions can be tested in a high-throughput expression platform. These
parameters include different baculovirus systems (Bac-to-Bac® and flashBACGOLD™)
with modified virus genome for enhanced secretion, signal peptides such as commonly
used honeybee melittin signal sequence and glycoprotein gp67 signal sequence,
insect cell lines (High Five™ and Sf9) and culture temperatures. Optimisation of the
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PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day One
upstream parameters is imperative for maximising protein yield for downstream
processing. To aid in recovery, recombinant glycoproteins are often expressed
with a widely used poly-histidine tag, which interacts with metal ions immobilised
on a chromatography matrix. However, purifying histidine-tagged glycoproteins
from spent culture supernatant presents a number of challenges. The presence of
interfering components in the cell culture media hinders direct loading of spent
culture supernatant onto chromatography column. Immobilised metal ions are
‘stripped’ from the purification media during sample loading, thus preventing the
capture of histidine-tagged proteins. Large volumes of spent culture supernatant and
endogenous host proteins can add further complexity to the recovery of secretory
glycoproteins. Here we present practical solutions for overcoming challenges
commonly encountered in expressing and purifying recombinant secretory
glycoproteins from the baculovirus-insect cell expression system.
Notes
Biography
Ms Cindy Chang is the Operations Manager of the Protein Expression Facility at the
University of Queensland. She has over 9 years of experience in recombinant protein
expression, ranging from project design, molecular cloning to microbial and animal
cell culture. Her diverse skill set in recombinant protein technology directs her in
developing well-thought-out strategies for projects, ensuring efficiency and costeffectiveness. With her in-depth understanding on the baculovirus-insect cell system,
she has implemented the high-throughput multi-hosts expression platform within
the facility. This streamlines the expression screen for multiple constructs with the
aim to determine optimal parameters for large-scale protein production.
Yeast expression systems
Brendon Monahan
CSIRO Manufacturing Flagship
Yeast have long served dual roles as research model systems and industrial
production platforms for a wide-range of biological activities and products.
Here I will present examples of utilising yeast as protein expression systems, in
both academic and industrial contexts. The presentation will focus on a number
of yeast species including the budding yeast Saccharomyces cerevisiae, and the
methylotrophic yeast Pichia pastoris (Komagataella phaffii). For each I will describe
the different strains and expression vector options available, as well as the respective
advantages and disadvantages. Finally, recent trends in using yeast as a tool for
the characterisation of small-molecule inhibitors of your target protein will also be
discussed.
Biography
Brendon Monahan is a research scientist in the Biosciences Program at CMSE,
project leader for the Cancer Therapeutics CRC, and a Lecturer in the Department
of Genetics, The University of Melbourne. Brendon is a yeast and fungal molecular
geneticist with a research background in epigenetics and gene regulation in various
fungal species. Over the last few years at CSIRO, Brendon has been involved in the
molecular genetic manipulation of yeast and fungal systems for the generation of
bio-based products, along with leading the protein production team for the Cancer
Therapeutics CRC.
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Mammalian toolbox: Platform technology for developing
human therapeutics
JEFF HOU
Australian Institute for Bioengineering and Nanotechnology
Biologics are the most rapidly growing class of human therapeutics, with the most
prominent class of biologics being monoclonal antibodies. Recombinant proteins are
manufactured through various host cell systems ranging from bacteria to yeast to
mammalian cells. The workhorse of the industry remains to be the Chinese Hamster
Ovary (CHO) cell; in fact over 70% of all approved human therapeutics worldwide is
manufactured in CHO cells.
Transient expression technology has gained significant interest in the field due to its
speed and flexibility in providing recombinant proteins for initial proof of concept
studies. This has significantly reduced timelines as well as gained further traction with
the emergence of follow-on biologics. Stable expression technology continues to push
the boundaries of mammalian cell culture with advances in host cell engineering and
bioprocess development pushing productivities to well above a couple of grams per litre.
At the National Biologics Facility (NBF), we use a high throughput platform process to
generate production cell lines for the manufacturing of recombinant proteins with a
focus on monoclonal antibodies. Novel molecules identified through discovery platforms
such as mAbLAB are reformatted and assessed using mammalian transient expression
technology such as EpiCHO followed by design and creation of a stably producing
mammalian cell line via either a ClonePix FL or flow cytometry facilitated cloning
method.
Apart from cell line engineering and platform design, NBF have large scale
manufacturing capabilities which includes a suite of equipment targeted at mammalian
cultivation, harvesting, purification and analytics to convert basic raw materials
into high-valued therapeutics. NBF’s current plant also uses a number of single-use
bioreactors and continuous culture as a mode of cultivation as a way to increase
productivity and reduce the environmental footprint.
Biography
Working with Prof. Peter Gray, A/Prof Stephen Mahler and Dr. Martina Jones, his part of
the key leadership group at the QLD Node to support advanced biologics manufacturing
in Australia. Dr. Hou’s background is in the area of mammalian host cell engineering
and adapting high throughput processing to the area of biologics manufacturing.
He has been involved in many industry projects taking lead candidates through CMC
development and developing a path into the clinic. His current role is the Operations
Manager at the National Biologics Facility (QLD Node) where his focused on developing
tools for manufacturing of biologics as well as delivering a manufacturing service to help
translate basic research.
16
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day One
Discovery and development of novel human therapeutics using
recombinant protein expression in mammalian cells
Notes
Catherine Owczarek
CSL Limited
Recombinant protein biologics such as monoclonal antibodies and coagulation
factors are becoming increasingly important in today’s pharmaceutical industry. As
they are secreted proteins with complex folding and post-translational modifications
they need to be generated in mammalian cells. Production of these proteins for preclinical studies using the technique of transient gene expression is an efficient, time
and cost-effective alternative to developing stable cell lines. Our current transient
gene expression methods enable us to produce mg-gram quantities of proteins
within 1-2 weeks from the time of generation of the appropriate protein expression
construct. We have evaluated the recently developed Expi293™ expression system
and compared it to our previously established platform that utilises the Freestyle
293™ system.
A combination of both transient expression technologies has allowed us to rapidly
screen, identify and characterise multiple novel protein-based human therapeutic
drug candidates.
Biography
Dr Catherine Owczarek leads the Recombinant Protein Expression Group located at
the Bio21 Institute. After gaining a PhD at the John Curtin School of Medical Research,
Canberra, Catherine completed her post-doctoral studies at the Sir William Dunn
School of Pathology in Oxford, the Walter and Eliza Hall Institute and then was a
Senior Research Fellow at the Monash Institute of Medical Research where she was
appointed a Senior Scientist of the Institute. Since joining CSL in 2004 Catherine has
directed the Research Group’s efforts in the successful development of a program to
produce mammalian-derived recombinant proteins in scaleable suspension culture
systems using disposable cell culture technology. She is involved in a range of early
phase drug discovery campaigns where there is a high demand for recombinant
proteins.
CareerLounge: Connecting students, employers and educators
Dominique Fisher
CareerLounge
CareerLounge Pty Ltd develops digital solutions for students, educators and
employers. CareerLounge was established in 2008 when co-founder, John Edward
Collins, discovered that there was a critical need for better communication between
students, educators and employers. CareerLounge offers a simple to use suite of
products designed to improve and enhance decision making, communications and
transactions between each of these related stakeholders.
www.comet.is and www.quillo.in have evolved from the original web application,
currently known as www.careerlounge.com.au, as two separate but highly
complementary products. Commercially launched in October 2012, www.
careerlounge.com.au is to be re-launched as Comet (www.comet.is) and Quillo
(www.quillo.in) in July 2014.
17
15th Annual Protein Expression Workshop
Day One
Notes
Comet is a global digital community made up of registered students, employers,
service providers and educators who create and use Comet Profiles (individuals) and
Comet Pages (organisations) to come together by industry sectors known as Comet
Villages. Each of the [43] Villages highlight opportunities available in that sector
by way of linked editorial and user generated content; and, relevant education and
employment opportunities in each Village.
Quillo, a complementary product, is the career pathway wiki organized by Villages that
provide users with formal industry definitions of job positions, career paths, preferred
education and training options and associated services.
Biography
Dominique is the Managing Director and Executive Chairman of CareerLounge Pty
Ltd (as well as one of its founders). For more than 30 years Dominique has worked
in communications telecommunications and information technology; specifically
in the application of technology in business and electronic commerce and the
commercialisation of new technologies. Dominique is a highly experienced executive,
and public and private company director and has held directorships with NRMA,
Insurance Australia Group (IAG), Pacific Brands, Sydney Opera House, The Malthouse
Theatre, Australia Council of the Arts, Dance Board, Prostate Cancer Foundation,
Royal Hospital for Women Advisory Board Chairman, AIDS Fund Raising Trust,
Communications and Media Law Assoc, was on the ICT Advisory Board to the Minister
for Communications, the Hon Helen Coonan, Chairman of Sky Technologies Pty Ltd,
and is Chairman of ASX listed Circadian Technologies Ltd.
18
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day TWO
Notes
19
15th Annual Protein Expression Workshop
Day TWO
Notes
Control of glycosylation profiles in cell bioprocesses
MICHAEL BUTLER
University of Manitoba
The glycoform profile of a monoclonal antibody (Mab) determines many functional
properties that affect therapeutic efficacy. Common variations of the conserved Fc
glycan include galactosylation, fucosylation and sialylation. The observed glycan profile
of the final product can depend upon the producer cell line, the growth media, the
culture conditions as well as the Mab protein structure. Post-translational modification
of proteins, particularly glycosylation is affected by culture conditions including the
availability of nutrients. During fed-batch and other cell culture strategies nutrients are
often maintained at low concentrations to ensure an efficiency of energy metabolism.
However, there is a possibility that these strategies may cause variability in the macroor micro-heterogeneity of glycosylation of synthesized glycoproteins. We have shown
that low nutrients levels present in fed-batch cultures may alter the glycan profile as
well as glycan occupancy by reducing the availability of the essential precursors during
glycosylation in the Golgi.
We have been able to produce Mabs with a wide range of glycan microheterogeneity in
culture by controlling the parameters of culture and by the use of several glycoprotein
processing inhibitors. The modified glycosylation profiles of the Mabs were determined
with HILIC (hydrophilic interaction liquid chromatography) analysis.
The presentation will review bioprocess parameters that can be controlled in order to
minimize batch to batch variation of Mab glycosylation. Strategies will also be discussed
to produce Mabs with pre-defined glycan structures.
Biography
Michael Butler is a Distinguished Professor at the University of Manitoba, Canada.
Previous appointments include Associate Dean of Research at the University of Manitoba
and Principal Lecturer in Biotechnology at Manchester Metropolitan University. He has
also been a Visiting Scientist at the Massachusetts Institute of Technology (USA), Animal
Virus Institue (Pirbright, UK) and the Universities of Oxford and Rio de Janeiro. He holds
degrees in Chemistry and Biochemistry from the Universities of Birmingham, London and
Waterloo (Canada).
His research focuses on the development of bioprocesses using mammalian cells in
culture for the production of recombinant proteins, monoclonal antibodies and viral
vaccines. He is particularly interested in the bioprocess conditions that can be used
to control the biochemical structure of glycoproteins. He has collaborated closely
with industry and in 2004 was the recipient of the Canadian national Synergy Award
for University-Industry innovation. He is presently scientific director of the Canadian
Network for Monoclonal antibody production (MabNet), which involves collaboration
between several universities and companies in Canada. He has published 7 books on
animal cell technology and >100 peer-reviewed scientific papers.
He is the founder and president of Biogro Technologies Inc., a spin-off company
dedicated to serum-free media development for mammalian cell bioprocesses. He is a
member of European Society Animal Cell Technology, Biochemical Society, Canadian
Society of Microbiology, Canadian Society Chemical Engineering and President of
the International Society for Protein Expression (PEACe). He is currently an editor of
Biotechnology Advances and on the editorial board of Biotechnology and Bioengineering.
20
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day TWO
Purification and characterisation of mammalian expressed
proteins: Key assessment considerations for candidate/target
selection in a research based commercial environment
Notes
pierre scotney and peter schmidt
CSL Limited
The discovery and development of drug candidates in the biopharmaceutical industry
involves the design, expression and purification of a large number of proteins such as
therapeutic antibodies. With custom gene synthesis and high-throughput sequencing
readily available to researchers, the bottleneck to the generation of larger numbers of
proteins has shifted to the purification of proteins in an efficient and economical way.
Therefore, despite the existence of well-established methods to purify antibodies in the
range of 1 – 100 mg (e.g. AKTA Xpress platform) there is a continuous need to further
miniaturise antibody purification in order to increase the throughput while reducing the
associated time and cost. In order to achieve these goals we have developed a highthroughput antibody purification platform suitable to purify up to 96 antibodies a day
in a time and cost-effective manner. The obtained yields are sufficient for biochemical
screening technologies. When the number of candidate molecules has been narrowed
down the production of drug candidates can be scaled up to obtain sufficient material
for in vitro and in vivo functional assays.
When screening libraries for drug candidates, such as therapeutic antibodies, the leads
are initially ranked by binding affinity to their target or potency in biochemical and cell
based assays. Biopharmaceuticals have characteristics associated with the molecule
being protein based which can affect the potential for the candidate to be developed into
a drug. We have implemented a screening process early in the drug discovery phase that
investigates the developability of lead candidate biopharmaceuticals by characterising
the protein’s biochemical and biophysical properties, such as post-translational
modification homogeneity, inherent and thermal stability, amenability to high
concentration formulation and particle analysis. The information from developability
profiling of candidate molecules early in drug discovery facilities in the selection of
which lead biopharmaceuticals will progress in the development process.
Biography: Pierre scotney
Dr Pierre Scotney is a Senior Scientist in Research at CSL Limited, as well as an
Honorary Member of the University Melbourne at the Bio21 Institute. He has extensive
experience in the pharmaceutical and biotechnology sector having worked at Roche
Pharmaceuticals, Zenyth Therapeutics and CSL Limited. He completed his PhD in
biochemistry at the University of Bristol (UK) in 1999 and came to Australia to undertake
postdoctoral research at St. Vincent’s Institute of Medical Research (Melbourne). In
2001 he began investigating the biology of Vascular Endothelial Growth Factors (VEGFs)
with an industry-partnered post-doctoral fellowship at the University of Melbourne and
biotechnology company Zenyth Therapeutics (then Amrad Corporation). In 2004 he
joined Zenyth to continue the research and develop novel therapeutics that targeted
VEGFs. In November 2006 Zenyth Therapeutics was acquired by CSL Limited and Dr
Scotney was appointed as a Senior Scientist, Research, located at the Bio21 Institute.
He currently works with a large multidisciplinary team focused on the discovery and
development of new recombinant antibody and protein-based medicines to treat
serious human disease. Dr Scotney has published in peer-reviewed publications and has
inventorship on patent filings.
21
15th Annual Protein Expression Workshop
Day TWO
Notes
Biography: peter schmidt
Peter started his scientific career at the Carl-von-Ossietzky University in Oldenburg,
Germany, in the department of Neuro-Biochemistry. In the following four years he
worked as PhD student at the Cardiovascular Research Centre of Bayer HealthCare in
Wuppertal, Germany, in the field of NO-mediated blood pressure regulation. Based on
this work, Peter was awarded an Alexander-von-Humboldt Fellowship which allowed
him to move to Melbourne and to continue his research for two more years at the
department of pharmacology of Monash University. After this short visit in academia
he started to work for four years at CSIRO in Parkville on the expression of human
influenza Neuraminidase. Since 3 years Peter is working as senior scientist in the Protein
Technologies group of CSL R&D in Parkville.
Structural biology at the Australian synchrotron
jose Varghese
CSIRO Manufacturing Flagship
Since first light in April 2007 the Australian Synchrotron at Clayton in Victoria has
ushered in a new era in biological research in Australia. Australian scientists have no
longer been limited by the availability of beamtime on a few overseas synchrotrons.
Over the last 7 years access to several different experimental platforms for research
into the structure of biological macromolecules have been providing insights into
structural biology. Two protein crystallography beamlines have been fully operational,
the first from a bending magnet and is designed to be a high throughput MAD (Multiple
Anomalous Diffraction) designed to exploit the use of seleno- methionine derivatives to
phase X-ray data from protein crystals. The other much more intense beamline from an
insertion device is designed to be able to collect diffraction data from crystals as small
as 10 microns in size. Other beamlines such as SAXS (Small Angle X-ray Scattering) for
determining the shapes of protein in solution, XAFS (X-ray Absorption Spectroscopy) for
studying metal protein interactions are currently available for users, and a CD (Circular
Dichroism) for studying protein secondary structure has been proposed.
Biography
Varghese was trained in Australia as a theoretical physicist with a background in
mathematics and natural philosophy. He did his post graduate training in experimental
physics and later in neutron physics. On returning to Australia he used his physical
science training to study biological systems, resulting in his multi-disciplinary approach
to solving problems in biology and acquiring skills in molecular and cellular biology.
This has enabled him to form and lead multidisciplinary teams of scientists that are
making inroads into complex biological problems. The impact of his work in influenza
is now being realized globally with emerging pandemics. Recently he was leading the
development of drugs and diagnostics for Alzheimer’s disease and mental disorders
in the P-Health Flagship. He has also acted in advisory roles for State and Federal
Governments in areas of scientific infrastructure in particular the Australian Synchrotron
where he led the construction of the two protein crystallography beamlines. He is
currently a CSIRO Retired Fellow.
22
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day TWO
Simple and low cost quality control of expressed proteins
by mass spectrometry
Notes
Nicholas A. Williamson
The Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne
Protein expression has become a key technology for biomedical research. The
integrity and quality of the final expressed product is often critical to the sucess of the
downstream experiments for which proteins were produced. Many people simply use
SDS PAGE analysis to check their expression products. This talk, however, will discuss the
inexpensive and straightforward use of mass spectrometry to explicitly determine the
primary structure of the expressed products. Examples will be shown that highlight the
ability of mass spectrometry to identify point mutations, truncations and modifications
in recombinant products that would not be detected by SDS PAGE. Examples of protein
complex formation and monitoring refolding experiments will also be shown. The key
point of this presentation is to show that mass analysis of expressed proteins is fast,
simple, cheap and can easily be made a routine quality control step for expressed protein
work.
Biography
Dr Nicholas Williamson is the manager of the Proteomics and Mass Spectrometry
Platform (MSPF) at the Bio21 Institute at the University of Melbourne. He has more than
20 years experience in proteomic mass spectrometry and is an expert in the primary and
secondary structure analysis of proteins and peptides. He now runs the Proteomics Core
facility which provides a unique, open-access mass spectrometry resource to research
groups located in Melbourne. The facility offers users highly subsidised direct access
to instrumentation and the expert training required to perform their own proteomic
research.
Physical techniques in a protein workflow
Shane Seabrook
CSIRO Manufacturing Flagship
There is no single method that
allows complete characterisation of
a bio-macromolecule and its various
interactions. The best insight is gained
when you can access as many techniques
as possible, making sure to develop a
robust understanding of your protein
and how it responds to its immediate
environment (its purification or storage
formulation) and with ligands of interest
(co-factors, drug candidates).
At CSIRO we routinely use thermal
stability and spectroscopic techniques
to help us figure out how a protein
behaves – by building multidimensional
insight we can better understand the impact the chemical and physical environment will
have on our target of interest. Most importantly, we clearly outline our expectations
from an assay; are we trying to develop a comprehensive understanding of the protein
23
15th Annual Protein Expression Workshop
Day TWO
Notes
behaviour, are we trying to make a protein more suitable for downstream analysis, or
are we screening for “hit” conditions, such as ligand binding assays and formulation
screening?
This seminar will summarize some of the techniques available and where they might
fit in to your experiment workflow, how you can make decisions based on the data
you’ve collected, and the alternative strategies that may be available.
Reference: Seabrook, S. A.; Newman, J., High-Throughput Thermal Scanning for Protein Stability:
Making a Good Technique More Robust. ACS Combinatorial Science 2013.
Biography
Shane completed his PhD (physical chemistry) at the University of Sydney, followed
by post-doctoral projects in France and the Netherlands on macromolecular and
biopolymer characterisation. Shane joined the biophysics in the CSIRO Materials
Science and Engineering division in late 2009, and has been working within the
Collaborative Crystallisation Centre (www.csiro.au/c3) developing methods that aid
structural biology. Shane is currently completing an MBA at the Melbourne Business
School.
Australian therapeutics pipeline
STEWART HAY
Therapeutic Innovation Australia
Therapeutic Innovation Australia is a not for profit organisation tasked with
implementing an Australian Government research infrastructure project. A key
objective for TIA with this project is to improve the efficiency and effectiveness
of translational health research in Australia. This presentation will briefly outline
TIAs research data gathered over the last twelve months and it will describe the
implementation of TIAs strategy to enable better translation of health discoveries
into clinical application.
TIA is, amongst its 18 investments, supporting protein production capabilities
through development of the National Biopharmaceutical Facility, a collaborative
between the University of Queensland and CSIRO.
Biography
Stewart Hay has a diverse background in the science industry having worked in
medical research, and the pharmaceutical, nutraceutical, biofuel and medical device
industries. He holds a PhD in the field of cancer cell biology and has conducted
research into diabetes, Langerhans Cell Histiocytosis, the characterisation of a novel
viral particle, antibody production and mechanisms of apoptosis. Over the last
decade Stewart has held various management roles leading to his appointment as
Chief Executive Officer at Therapeutic Innovation Australia. In this capacity Stewart
manages the provision of funding for development of translational research facilities
and several national advisory committees.
24
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day TWO
The National Biologics Facility (NBF) – A joint AIBN/CSIRO facility
Notes
PETER GRAY
Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland
Biologics are a class of human therapeutics comprised of proteins such as monoclonal
antibodies produced by recombinant DNA technology. The specificity and other
advantages of biologics has meant that they have rapidly gained acceptance, eg: seven
out of the world’s top ten selling drugs are now biologics; biologics now account for 17%
of the total sales of pharmaceuticals; and annual sales of biologics are growing at 20%
per annum, compared to 7% for traditional small molecule pharmaceuticals.
For the above reasons, the NCRIS (National Collaborative Research Infrastructure
Scheme) Roadmap of February 2006 provided funding to establish national research
infrastructure in the area of Biologics. Following a review in 2010, this national research
infrastructure was consolidated into the ‘National Biologics Facility’ (NBF) consisting of
two nodes, one located at the AIBN in Brisbane, and the other located at the CSIRO in
Melbourne. Since 2006, the two nodes of the NBF have attracted total funding of over
$28 million from users, NCRIS (2006), EIF/SSI, NCRIS (2013) TIA, Queensland and Victorian
State Governments, and CSIRO and AIBN/UQ sources.
Currently NBF has 22 staff working in the two nodes. The NBF has gained an excellent
reputation with the Australian research community for the high quality of its work and
services. Over the last five years the NBF has completed over 500 projects for a number
of SME’s and public sector research organisations, and has produced and purified over
10,000 litres of mammalian cell cultures, and over a kilogram of purified protein. The
projects carried out include mammalian cell-line development, bioprocess development,
monoclonal antibody and vaccine isolation and production, and the production of preclinical amounts of biologics for animal and other testing.
Biography
Professor Peter Gray was appointed the inaugural director of the Australian Institute of
Bioengineering and Nanotechnology (AIBN) at the University of Queensland in 2003.
Previously he was Professor and Head of Biotechnology at UNSW and Director of the
Bioengineering Centre, and a Senior Principal Research Fellow at the Garvan Institute of
Medical Research in Sydney.
Professor Gray has had commercial experience in the USA working for Eli Lilly and Co and
for the Cetus Corporation as well as previously holding academic positions at University
College London, and at the University of California, Berkeley.
His research interests are focussed on engineering mammalian cells to produce the
complex proteins called biologics which are gaining rapid acceptance as human
therapeutics, and on developing human stem cells bioprocesses suitable for clinical
application.
Professor Gray was one of the founders and is a past President of the Australian
Biotechnology Association, AusBiotech. He is a Fellow of the Australian Academy
of Technological Sciences and Engineering (ATSE) and of the Australian Institute of
Company Directors, and has been named as one of Australia’s 100 Most Influential
Engineers. Awarded the Centenary Medal by the Australian Government in 2001.
He is a Vice-President of ATSE, and serves on the Boards of Biopharmaceuticals Australia
Pty Ltd, ACYTE Biotechnology Pty Ltd, Stem Cells Ltd, ECI Inc, New York, and a number of
State and Federal Government Councils and Committees.
25
15th Annual Protein Expression Workshop
Day TWO
Notes
BPA and the biologics facility of the future
BRETT WHITECROSS
BioPharmaceuticals Australia
BioPharmaceuticals Australia (BPA) was established to support the country¹s
biopharmaceutical development industry and fill a critical national capability gap
identified by both the Commonwealth and QLD State Governments ­GMP mammalian cell
product contract manufacturing. In October 2013, as a result of collaboration between
Government, industry, philanthropy, and the research sector, a $65M biologics facility
was opened in Brisbane. A partnership between BPA and Patheon Biologics (formerly
DSM Biologics) has seen the multinational CMO establish a subsidiary in Australia and
operate the world-class ³biologics-facility-of-the-future².
BPA continues to support this collaboration and strengthen the local industry through
the Biopharmaceutical Development Fund (BDF) ­designed to assist not-for-profit
organisations and fledgling biotech companies access this new national capability, while
attracting new developers to Australia.
Biography
Brett Whitecross has 8 years¹ experience in the biotechnology sector, working at the
interface between the public and private sectors and in a variety of capacities. As
Business Development Manager for BioPharmaceuticals Australia (BPA) Brett has been
involved in the establishment of a world-class biologics manufacturing facility for
mammalian cell products, the attraction of a multinational Contract Manufacturing
Organisation to Australia as BPA¹s operating partner, and biopharma industry
development. Brett is continuing with BPA¹s strategic initiatives, working with a global
focus to attract clients to the new facility.
Previously, Brett has held business development positions with PharmaSynth and
AusBiotech, has worked in policy development for the Government, and currently serves
as a committee member of AusBiotech¹s Queensland Branch.
Brett has received a Bachelor of Biotechnology Innovation with honours from the
Queensland University of Technology (QUT) and an MBA from the University of
Queensland (UQ).
The Victorian Antibody Initiative (VAI)
Caroline Laverty
Monash Antibody Technologies Facility
A joint initiative between the Walter and Eliza Hall Institute (WEHI) and Monash Antibody
Technologies Facility (MATF) now provides the biomedical research community with an
antibody partner fully attuned to the demands of world-class science.
The Victorian Antibody Initiative (VAI) brings together the state-of-the-art automated
production facilities at MATF with the premier customised development services at WEHI
for the very best in monoclonal and polyclonal antibody production.
For more than two decades, the Walter and Eliza Hall Institute’s Antibody Facility has
been integral to the institute’s leading position in biomedical research and developing
26
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day TWO
pre-clinical drug candidates and has earned a reputation for providing world-class
antibody development services. The personalised approach to customer service
guarantees that academic and industry users return to this facility time and again for
their antibody requirements.
Notes
In 2008, investment by Monash University, the Victorian State Government and the
Australian Federal Government (Bioplatforms Australia) created scale through the
Monash Antibody Technologies Facility, one of the most sophisticated, high-throughput
monoclonal antibody facilities in the world.
In partnership, two of Australia’s leading and complementing antibody facilities can
provide you with a “one stop” point of contact to access the best technology and
customer service for production of your immunological tools and reagents.
The Victorian Antibody Initiative is lead by a team having exceptional experience in
antibody design and production, including Kaye Wycherley and Paul Masendycz from
The Walter and Eliza Hall Institute Mark Sleeman and Caroline Laverty from Monash
University.
Biography
I studied at University College London (UCL) to obtain my PhD in physical-organic
chemistry under Prof. Mike Abraham. After which I moved to Pfizer as a Team Leader in
the Drug Metabolism Automation Team where we were responsible for the development
of a fully automated high throughput screening system for thousands of novel drug
compounds. As part of an exciting life change, I moved to Australia and joined Monash
University. Firstly at the Center for Drug Candidate Optimisation for 2 years and then
the challenge of getting a fully automated facility for the production of monoclonal
antibodies up and running brought me to MATF in 2009.
GMP Considerations for recombinant protein manufacture
and working with a CMO
BRETT BUTCHER
PharmaSynth
TRecombinant proteins can be produced in a diverse array of microbial and mammalian
cell lines. There are many considerations and decisions to be made as you take your
recombinant protein from the laboratory benchtop into the clinic including developing
manufacturing processes that are amenable to cGMP, include an element of future
proofing and produce the optimum yield to ensure cost effective manufacturing. Once
you have made the decision to outsource your manufacturing, the importance of
selecting the right CMO cannot be understated as:
•Biologics manufacturing is a technologically complex, highly regulated process.
•Large complex protein structures are unstable and have a low tolerance for error.
•Even the best product can fail if it experiences substantial delays in the
development process.
•There is a time commitment involved in transferring the technology into the
CMO required to make your recombinant protein.
27
15th Annual Protein Expression Workshop
Day TWO
Notes
Biography
Brett Butcher has worked for Progen Pharmaceuticals since 2002, performing the role
of Senior Scientist, specialising in the manufacturing and downstream processing of
pharmaceuticals. In 2009 he became the Production Manager of PharmaSynth and
coordinated Progen’s anti cancer drug PI-88s progression from preclinical to phase 3
manufacture. He has also been responsible for the tech transfer and manufacture of
many client products for both clinical applications and marketed products. Brett holds a
PhD in yeast biochemistry which investigated real time membrane fluidity modulation as
part of the adaptative response of yeast.
Insulin receptor: Production of midi-, mini- and micro-constructs:
Targeted design of insulin receptor for crystallography of Site 1
John Menting
Walter and Eliza Hall Institute of Medical Research
Investigating the structural biology of the human insulin receptor brings challenges
both in terms of protein production and in terms of crystallography. We have utilized
two constructs of the human insulin receptor using stably-transfected CHO Lec8 cells.
The first produces a thrombin-cleavable form of the three-domain L1-CR-L2 described as
a “minireceptor”. Thrombin cleavage yields the two domain L1-CR “micro-receptor” that
is capable of binding insulin in the presence of the receptor’s so-called alphaCT peptide.
A further construct, consisting of a homodimer of a L1-CR-L2-(FnIII-1)-alphaCT receptor
polypeptide (a “midi-receptor”) was produced and purified by insulin-affinity and
size-exclusion chromatography. The micro- and midi-receptor constructs yield crystals
in CSIRO C3 screens and their structures have been solved by molecular replacement.
Despite crystallographic limitations, a consequence of protein heterogeneity, these
structures are the first to reveal the way in which insulin binds to its primary receptor
binding site and the manner in which the insulin B-chain C-terminal segment is displaced
from the hormone core upon receptor engagement.
Biography
John Menting completed a B.Sc.(Hons) at the University of Melbourne in 1986 with
Professor Gerhard Schreiber studying the transport of thyroid hormones across the
blood-brain barrier. He completed a Ph.D. at La Trobe University with Professor Bob
Scopes working on a plant cytochrome P450 related to the “blue rose” project of Calgene
Pacific (now Florigene) in 1993 and was a member of the team that isolated and patented
the “blue gene”.
In two postdoctoral fellowships, first at WEHI with Professor Alan Cowman and then
at the Monash Microbiology Department with Professor Ross Coppel, he studied drug
resistance in malaria and the merozoite surface protein MSP4. He then joined the
Structural Biology Division at WEHI and worked with Dr. Jacqui Gulbis on translocases
of the mitochondrial outer membrane until in 2008 he changed course and established
the laboratory of Associate Professor Mike Lawrence shortly after Mike moved to WEHI
from the CSIRO at Parkville. Since then he has continued the investigation of the insulin
receptor and insulin-like growth factor receptor 1 leading to the publication in 2013 of
the first structures of insulin bound to Site 1 of the insulin receptor.
28
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day TWO
Telomerase: A promising target for cancer
Notes
Scott Cohen
Children’s Medical Research Institute
Telomeres, the repetitive DNA-protein complexes at the ends of linear chromosomes,
shorten with each cycle of DNA replication, providing a counting mechanism to limit the
number of times a cell can divide. Most cancer cells have activated the ribonucleoprotein
enzyme telomerase to add telomeric DNA repeats and thereby counteract telomere
shortening, allowing for unlimited proliferation; in contrast, normal cells have
undetectable or low levels of telomerase. Inhibition of telomerase is therefore a
promising avenue for future anticancer therapy that should be effective against a broad
range of cancers while displaying few side effects.
This talk will begin with a brief overview of telomere and telomerase biology, and
then cover the journey from an enzyme complex of unknown composition through
its purification, identification of components, and development of an overexpression
system – research that from the outset has involved collaboration with the Fermentation
Lab at CSIRO Materials Science & Engineering. The presentation will conclude with
progress towards a low-resolution EM structure of the human telomerase enzyme
complex.
Biography
Dr. Scott Cohen received a PhD in Chemistry from the California Institute of Technology,
elucidating the molecular mechanism of the “enediyne” class of naturally-occurring
DNA-cleaving agents. From Caltech he moved to the laboratory of Professor Tom Cech
at the University of Colorado to study the mechanism and conformational dynamics of
large catalytic RNA molecules (ribozymes). In 2003 he began his research into telomerase
at Children’s Medical Research Institute (Westmead NSW), and in 2007 established the
composition of the human telomerase enzyme complex, reported in Science. Dr Cohen’s
singular research aim is telomerase structure.
29
15th Annual Protein Expression Workshop
Day THREE – HANDS ON
Workshops
30
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Day TWO
Notes
31
15th Annual Protein Expression Workshop
Participants List
DurgaAcharya CSIRO
durga.acharya@csiro.au
Campbell
Aitken
CSL Limited
cheryl.whiteley@csl.com.au
Layla
Alhasan
RMIT University
s3314715@student.rmit.edu.au
HameedaAl-Musawi BMS
hameeda.al-musawi@bms.com
Jess AndradeCSIRO
jessica.andrade@csiro.au
Main Ern
Ang
University of Adelaide
main.ang@student.adelaide.edu.au
David
Antonjuk
BioNovus Life Sciences
david.a@bionovuslifesciences.com.au
Thilini
Anupama
University of Melbourne
nanupama@student.unimelb.edu.au
Tracee
Archibald
GE Healthcare Life Sciences
Michelle
Audsley
Monash University
michelle.audsley@monash.edu
Samar
Babkair
Burnet Instiute
samar.babkair@burnet.edu.au
Matt
Banfield
Sigma Aldrich
matt.banfield@sial.com
Paul
Barrett
ATA Scientific
pbarrett@atascientific.com.au
Sue BarrettCSIRO
susan.barrett@csiro.au
Rebekah
rebekah.bernard@rmit.edu.au
Bernard
RMIT University
KanikaBhadoria Sartorius
Kanika.Bhadoria@sartorius-stedim.com
Elham
Bidram
University of Melbourne
elhamb@student.unimelb.edu.au
Jamie
Black
CSL Limited
cheryl.whiteley@csl.com.au
Melissa
Blues
Merck Millipore
melissa.blues@merckgroup.com
Violeta
BogdanoskaEppendorf
MarkBourke Labtek
mark@labtek.com.au
Kerry BreheneyCSIRO
kerry.breheney@csiro.au
Tracy
Bryan
Children’s Medical Research
Institute
tbryan@cmri.org.au
Ashlee
Burnet Institute
ambur11@student.monash.edu
Burt
BrettButcherPharmaSynth
brett.butcher@pharmasynth.com.au
RebeccaButcher
CSL
rebecca.butcher@csl.com.au
Michael
Butler
University of Manitoba
butler@cc.umanitoba.ca
Avinesh
Byreddy
Deakin University
arbyredd@deakin.edu.au
Steven
Byrne
CSL Limited
cheryl.whiteley@csl.com.au
Teresa
Carvalho
Monash University
teresa.carvalho@monash.edu
Cindy
Chang
The University of Queensland
c.chang2@uq.edu.au
Zhian
Chen
Monash University
zhian.chen@monash.edu
Chris
Chiu
Walter and Eliza Hall Institute
of Medical Research
chiu@wehi.edu.au
Jocelyn
Choo
Monash University
mfcho2@student.monash.edu
Seong Hoong
Chow
Monash University
seong.chow@monash.edu
Children’s Medical Research
Institute
scohen@cmri.org.au
Scott
Cohen
PeterCollins CareerLounge
nikki.rees@careerlounge.com.au
Richard
Cormick
3M Purification
rcormick@mmm.com
David
Cossens
GE Healthcare Life Sciences
David.Cossens@ge.com
HelenDacres CSIRO
helen.dacres@csiro.au
Emma
Dal Maso
Monash University
emma.dalmaso@monash.edu
Greg
D’Cruz
Box Hill Institute
g.d’cruz@boxhill.edu.au
Stephen
de Lacey
Eppendorf
deLacy.S@eppendorf.com.au
Matthew
Dennis
Monash University
matthew.dennis@monash.edu
Pathum DhanapalaCSIRO
32
bogdanoska.v@eppendorf.com.au
pathum.dhanapala@csiro.au
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
Alison Digney
Life Technologies
Alison.Digney@thermofisher.com
Esin
Donaldson
CSL Limited
esin.donaldson@csl.com.au
TheresaDowney
CSIRO
theresa.downey@gmail.com
Valentina Patrys Limited
vdubljevic@patrys.com
Dubljevic Geoff DumsdayCSIRO
geoff.dumsday@csiro.au
Sarah
sarah.edwards@csiro.au
Edwards
Monash University & CSIRO AAHL
Heidi EL-TahhanCSL
heidi.el-tahhan@cslbehring.com.au
RochelleEnright
Genesearch
rochellee@genesearch.com.au
Lou CSL
Louis.Fabri@csl.com.au
FriederikeFehr
CSIRO
friederike.fehr@csiro.au
DominiqueFisher
CareerLounge
nikki.rees@careerlounge.com.au
Chwan-HongFoo
CSIRO
chwan.foo@csiro.au
Mike
Circadian Technologies Ltd
mike.gerometta@circadian.com.au
Fabri
Gerometta
VeronicaGlattauer CSIRO
veronica.glattauer@csiro.au
Benjamin
Gloria
Ludwig Institute for
Cancer Research
benjamin.gloria@ludwig.edu.au
Christopher
University of Melbourne
c.gonelli@student.unimelb.edu.au
Gonelli
Julian GrusovinCSIRO
julian.grusovin@csiro.au
Jun
Burnet Institute
jun.gu@burnet.edu.au
Christoph
Hagemeyer
Baker IDI Heart and Diabetes
Institute
christoph.hagemeyer@gmail.com
Joshua
Gu
Monash University
joshua.m.hardy@monash.edu
NatalieHayes
Hardy
Eppendorf
shir.n@eppendorf.com.au
ThomasHayman
WEHI
tomhayman001@hotmail.com
Jack HertzogMonash
jack.hertzog@monash.edu
Behnaz
University of Melbourne
behnazh@student.unimelb.edu.au
AnitaHill
Heydarchi
CSIRO
anita.hill@csiro.au
Jeremy
Hoglin
VWR International
jeremy.hoglin@au.vwr.com
Jeff
Hou
University of Queensland, AIBN
jeff.hou@uq.edu.au
LindaHowell CSIRO
linda.howell@csiro.au
Jon
Ince
GE Healthcare Life Sciences
jon.ince@ge.com
Amelia
Jalanski
CSL Limited
cheryl.whiteley@csl.com.au
Edwina
Jap
Baker IDI
edwina.jap@bakeridi.edu.au
AnthonyJay
PharmaSynth
anthony.jay@pharmasynth.com.au
Carina
RMIT University
carina_ joe@y7mail.com
MariettaJohn
Joe
CSIRO
marietta.john@csiro.au
Martina
Jones
The University of Queensland
martina.jones@uq.edu.au
Michael
Keir
Lonza Australia Pty Ltd
michael.keir@lonza.com
Sadia Nawroz
Khan
Monash University
sadia.khan@monash.edu
KeithKhoo
CSIRO/Deakin
keith.khoo@deakin.edu.au
Tum
Khwanmuang
ENFA Pty Ltd
tum@isciencetech.com.au
Natalie
Kikidopoulos
RMIT University
natalie.kikidopoulos@rmit.edu.au
Anastassija
Konash
Universal Biosensors
skonash@universalbiosensors.com
Mellisa
Kowalski
GE Healthcare Life Sciences
MylinhLa
Samantha
Lagaida
CSIRO
mylinh.la@csiro.au
University of Melbourne
slagaida@student.unimelb.edu.au
CarolineLaverty
MATF
caroline.laverty@monash.edu
Rachel
Monash University
rachel.lee@monash.edu
Lee
33
15th Annual Protein Expression Workshop
Jo LevettSartorius
jo.levett@sartorius.com
Yulia
Losev
Baker IDI Heart and Diabetes
Institute
yulia.losev@bakeridi.edu.au
GeorgeLovrecz
CSIRO
george.lovrecz@csiro.au
LouisLu
CSIRO
louis.lu@csiro.au
JasminaLuczo
CSIRO
jasmina.luczo@csiro.au
Therese
Lynch
CSL Limited
therese.lynch@csl.com.au
Siti Nurfatimah
M.Shahpudin
Monash University
sitinurfatimah02@gmail.com
Mayra
Machuca
Monash University
mayra.machuca.perez@monash.edu
Luis Malaver
Malaver Ortega
CSIRO
luis.malaver@csiro.au
Kirstie
Mangas
University of Melbourne
kmangas@student.unimelb.edu.au
Ashley
Marsh
Murdoch Childrens Research
Institute
ashley.marsh@mcri.edu.au
Barry
Marshall
University of Western Austrlia
klaurie@helicobacter.com
Kelly
McDonaghGRACE
Kelly.McDonagh@grace.com
Bill
McKinstryCSIRO
bill.mckinstry@csiro.au
James
McRae
3M Purification
jrmcrae@mmm.com
Bronwyn
Meehan
University of Melbourne
bmeehan@unimelb.edu.au
John
Menting
Walter and Eliza Hall Institute
for Medical Research
menting@wehi.edu.au
Daryn
Metti
Bioscientific pty ltd
daryn@biosci.com.au
Ann
Mills
POCD Scientific
amills@pocd.com.au
OskarMills
Private
s3337250@student.rmit.edu.au
Joyanta Kumer
Monash University
joyanta.modak@monash.edu
Modak
BrendonMonahan CSIRO
brendon.monahan@csiro.au
Naomi
Life Technologies
Naomi.Morison@thermofisher.com
Jacqueline
Morris
Morison
CSIRO Australian Animal Health
Laboratory
jacqui.morris@csiro.au
Bevan
Thermo Fisher Scientific
bevan.morton@thermofisher.com
Morton
Matthias NachtschattCSIRO
matthias.nachtschatt@csiro.au
Musammat
University of Melbourne
musammat.nahar@student.unimelb.edu.au
NishenNaidoo
Nahar
Bio-rad
nishen_naidoo@bio-rad.com
Amy
Nankervis
Pall Australia
amy_nankervis@ap.pall.com
Vani
Narasimhulu
Burnet Institute
vani@burnet.edu.au
Hans
Netter
Monash University
hans.netter@monash.edu.au
Aaron
Ng
3M Purification
ang2@mmm.com
Liza
Nguyen
Bristol-Myers Squibb
liza.nguyen@bms.com
JulieNigro CSIRO
julie.nigro@csiro.au
Amir
Noormohammadi
University of Melbourne
amirh@unimelb.edu.au
Gail
O’Connell
In Vitro Technologies
marketing@invitro.com.au
Catherine
Osborne
Thermo Fisher Scientific
catherine.osborne@thermofisher.com
Catherine
Owczarek
CSL Limited
catherine.owczarek@csl.com.au
Lorien
Parker
St. Vincents Institute
lparker@svi.edu.au
PareshParmar
CSIRO
paresh.parmar@csiro.au
Timothy
Patton
Burnet Institute
timothy.patton@burnet.edu.au
Hannah
Pearce
Baker Heart Research Institute
hannah.pearce@bakeridi.edu.au
CSIRO
lesley.pearce@csiro.au
LesleyPearce
34
PROTEINS FOR STRUCTURAL AND BIOLOGICAL STUDIES
GrantPeck
CSIRO
grant.peck@csiro.au
YongPeng
CSIRO
yong.peng@csiro.au
AngeloPerani
LICR
angelo.perani@ludwig.edu.au
Jessica
CSL Limited
jessica.petracca@csl.com.au
Petracca
TamPham CSIRO
TramPhan
CSIRO
tam.pham@csiro.au
tram.phan@csiro.au
Pat PillingCSIRO
pat.pilling@csiro.au
Luisa
luisa.pontes-braz@csiro.au
Pontes-BrazCSIRO
JOHNPOWER ZOETIS
john.power@zoetis.com
Rasika
Premaratne
Monash University
dinamithra.premaratne@monash.edu
Tam
Quach
CSL Behring
tam.quach@cslbehring.com.au
Adam
Quek
Monash University
juinn.quek@monash.edu
Rebecca
Rabi
Monash University
rebecca.rabi@monash.edu
Soizic RadulovicCSIRO
soizic.radulovic@csiro.au
AnnaRaicevicCSIRO
anna.raicevic@csiro.au
John RamshawCSIRO
john.ramshaw@csiro.au
Ali
Razaghi
James Cook University, Australia
ali.razaghi@my.jcu.edu.au
Peter
Regan
Kinesis Australia
peterr@kinesis-australia.com.au
Howard
Roberts
Millennium Science
HRoberts@mscience.com.au
Chiara
Ruggeri
Monash university
ruggeri.mena@gmail.com
Rajendra
Sabale
Serum Institute of India Ltd
rajendra.sabale@seruminstitute.com
Abu Iftiaf Md.
Salah Ud-Din
Monash University
abu.ud-din@monash.edu
Bansi
Sanghvi
Bio-Rad Laboratories
bansi_sanghvi@bio-rad.com
PaulSavageCSIRO
paul.savage@csiro.au
PeterSchmidtCSL
peter.schmidt@csl.com.au
JudyScoble CSIRO
judy.scoble@csiro.au
Pierre
pierre.scotney@csl.com.au
Scotney
CSL Limited
Shane SeabrookCSIRO
shane.seabrook@csiro.au
Michiel
TrendBio Pty Ltd
michiels@trendbio.com.au
KayleneSelleck
CSIRO
kaylene.selleck@csiro.au
Madhu Sharma
Monash University
madhu.sharma@monash.edu
BrianShiell
CSIRO
brian.shiell@csiro.au
sean
Omni Executive
sean.smith@omniexe.com
A&D Australia C/- Labtek
tims@andaustralasia.com.au
Knoesis Biotech
vishal@knoesisbiotech.com.au
Selen
smith
Tim
Vishal
Sengupta
Smith
Srivastava
Stef
StefanakosGRACE
Stef.Stefanakos@grace.com
Randy
SuryadinataCSIRO
randy.suryadinata@csiro.au
Yuzhe
Tang
yuzhe.tang@adelaide.edu.au
University of Adelaide
Han TanwarCSIRO
han.tanwar@csiro.au
Nick
VWR International
nick.tartaris@au.vwr.com
Anais
Tatry
Tartaris
Ludwig Institute for
Cancer Research
anaistatry@hotmail.fr
Jan
Tennent
Biomedical Research Victoria
jan.tennent@biomedvic.org.au
Stephen
Tibb
3M Purification
sjtibb@mmm.com
Tak
Tiong
Assay Matrix Pty Ltd
tak@assaymatrix.com
Mary
Triantis
bioscientific pty ltd
mary@biosci.com.au
35
15th Annual Protein Expression Workshop
NickTsichlisLonza
nick.tsichlis@lonza.com
Jaana
Tuominen
Walter and Eliza Hall Institute
tuominen.j@wehi.edu.au
Birgit
Unterweger
Monash University/CSIRO
birgit.unterweger@monash.edu
Jose
VargheseCSIRO
GRACE
Gina.Vello@grace.com
Trent Warburton
TrendBio Pty Ltd
trentw@trendbio.com.au
Andri
Wardiana
The University of Queensland
andri.wardiana@uq.net.au
Thomas
Watts
Monash University
thomas.watts@monash.edu
Christopher
Weir
Walter and Eliza Hall Institute
weir.c@wehi.edu.au
Jerome
WerkmeisterCSIRO
Brett
Whitecross
jerome.werkmeister@csiro.au
BioPharmaceuticals Australia (BPA) brett.whitecross@biopharmaus.com.au
Eden WhitlockCSIRO
eden.whitlock@csiro.au
DeborahWilliams
Pall
deborah_williams@ap.pall.com
Nicholas
University of Melbourne
nawill@unimelb.edu.au
Williamson
TomWilsonCSIRO
tom.wilson@csiro.au
Elton
Wong
Monash Institute of
Pharmaceutical Sciences
elton.wong@monash.edu
Janice
Woods
Millennium Science
jwoods@mscience.com.au
Amanda
Woon
Monash University
amanda.woon@monash.edu
Guojie
Wu
Monash University
wuguojie@sina.cn
CSIRO
xiaowen.xiao@csiro.au
XiaowenXiao
36
jose.varghese@csiro.au
GinaVello
Li
Yang
Monash University
li.yang@monash.edu
Wei
Yang
Monash University
wei.yang@monash.edu
Kuok
Yap
ARC Centre of Excellence in
Plant Cell Walls
kuok.yap@adelaide.edu.au
Mike
Yarski
Millennium Science Pty Ltd
myarski@mscience.com.au
Bonnibel
Yee
Genesearch Pty Ltd
bonniyee@genesearch.com.au
RimaYouil
Private
rimayou@yahoo.com
Ling
Zhu
University of Melbourne
lingz1@student.unimelb.edu.au
Rick
Zolzinski
Merck Millipore
rick.zolcinski@merckgroup.com
NOTES
MANUFACTURING FLAGSHIP
www.csiro.au
Recombinant Protein
Production Facility
CSIRO is Australia’s national science agency. We employ 6500 people and are ranked in the
top 1% of research organisations in the world. Our inventions include the Relenza flu treatment
and fast WLAN. Our Recombinant Protein Production Facility (RPPF) is an important part of our
research capability – helping to facilitate life-changing research across the globe.
Dr George Lovrecz at
CSIRO’s Recombinant
Protein Production Facility.
About the RPPF
The RPPF provides access to stateof-the-art laboratories and expertise
relating to the production, processing,
development and optimisation of
recombinant proteins. The facility is
based in Melbourne and is supported by
Australia’s Super Science Initiative.
One of the challenges of bringing
research and development to a
commercial reality is the production
of potentially valuable proteins in
quantities large enough to support precommercial investigations and trials.
The RPPF is supported by the expert
capabilities of various molecular and
cell biology groups. We can facilitate
the optimisation, scaling-up, production
and purification of recombinant proteins
in large quantities: from hundreds of
milligrams to kilograms.
Our expertise
Our capabilities include a proven track
record in fermentation and downstream
processing of recombinant proteins,
expressed in eukaryotic and prokaryotic
hosts, such as mammalian, insect, yeast
and bacterial cells.
Recent projects included the:
◆ production of monoclonal antibodies
◆ receptor, signalling and cytokine
proteins
◆ bacterial and insect cell proteins
◆ various enzymes
◆ food additivies.
The RPPF provides national access to
process development, optimisation and
protein production in line with bestpractice standards including ISO9011
certification for tissue culture work.
Case Study: Antibodies for
developing countries
In 2012 CSIRO was contracted by
Program for Appropriate Technology
in Health (PATH), an international
not-for-profit organisation, to
produce antibodies on a large scale
to aid the development of new, safe,
affordable and effective vaccines
against rotavirus.
Rotavirus kills nearly half a million
children every year, 85 per cent of
whom live in developing countries.
PATH is working to increase access
to and effectiveness of existing
commercial rotavirus vaccines
worldwide.
CSIRO used their recombinant
protein production facility for the
large-scale ISO9001 production
of six antibodies that will be used
to test the quality of the rotavirus
vaccines being developed.
“This unique, state-of-the-art facility
allowed us to produce enough
antibodies to supply PATH’s partners
for the next five years,” said Prof
Lovrecz.
The antibodies, developed by
international researchers including
the Murdoch Children’s Research
Institute, will be used by PATH’s
manufacturing partners in China,
Brazil, and India and by the United
States’ National Institutes of Health.
Researchers controlling CSIRO’s
Brunswick reactor.
Dr Louis Lu working in CSIRO’s Recombinant Protein Production Facility.
Facilities
The state-of-the-art fermentation
laboratories are equipped with a wide
range of cell culture and fermentation
equipment including:
◆ and large-scale protein production
◆ single use bioreactors up to 25L
working volumes
◆ roller bottle apparatus, spinner and
shaker flasks suitable for scale-up and
process development work
◆ a wide variety of analytical equipment
to follow cell growth and metabolism
and characterise proteins
◆ downstream and purification
equipment suitable to process up 500L
batches of bacterial and mammalian
cell culture harvests.
cell biology groups to allow cell line
development, protein chemistry for
rapid purification and characterisation
of proteins. Our direct connection to the
C3 crystallisation laboratories and to the
Australian Synchrotron to also allows
structural studies.
Collaboration and funding
The Australian Government, State
Government of Victoria and CSIRO
have invested in this facility to enhance
Australia’s capacity to produce
recombinant proteins in pre-commercial
quantities.
The RPPF closely collaborates with The
University of Queensland’s Australian
Institute for Bioengineering and
Nanotechnology, the Monash and WEHI
Monoclonal Antibody Facilities.
The RPPF is supported by the expert
capabilities of various molecular and
CONTACT US
YOUR CSIRO
FOR FURTHER INFORMATION
t
Australia is founding its future on
science and innovation. Its national
science agency, CSIRO, is a powerhouse
of ideas, technologies and skills for
building prosperity, growth, health and
sustainability. It serves governments,
industries, business and communities
across the nation.
Manufacturing Flagship
Prof George Lovrecz
t +61 3 9662 7348
e george.lovrecz@csiro.au
w www.csiro.au
1300 363 400
+61 3 9545 2176
e enquiries@csiro.au
w www.csiro.au
Contact us
t 1300 363 400
+61 3 9545 2176
eenquiries@csiro.au
wwww.csiro.au
Your CSIRO
Australia is founding its future on
science and innovation. Its national
science agency, CSIRO, is a powerhouse
of ideas, technologies and skills for
building prosperity, growth, health and
sustainability. It serves governments,
industries, business and communities
across the nation.