WHERE DOES YOUR MONEY GO ?
Your contributions make a very real difference to our research
EQUIPMENT
State-of-the-art spectrophotometer
[www.bio-logic.info/instruments/mos-500]
Amyloidosis is caused by the formation of abnormal amyloid fibril deposits
within the body. One of the unsolved mysteries of amyloidosis is the question
of how these fibrils all have similar structures, regardless of which of the
original, highly varied “precursor” proteins they are formed from. The
Wolfson Drug Discovery Unit has recently acquired a new, state-of-the-art,
highly sophisticated piece of equipment, a MOS-500 fast modular
spectrometer/polarimeter, from Bio-Logic. Researchers believe this will help
to improve our understanding of this conundrum.
Dr Patrizia Mangione
The new spectrometer was purchased with funds from the ARF and it is
already proving invaluable in helping us unravel the processes whereby
different “globular” proteins unfold in the early stages of their journey to
becoming amyloid fibrils.
Dr Patrizia Mangione, working in Professor Vittorio Bellotti’s Protein Misfolding Group, is shown using this
new instrument. Until recently she had to travel to Pavia in Italy to have access to this technology. Patrizia
says that the new insights into amyloid fibril formation gained from our new equipment will help to bring us a
step closer to understanding how amyloid fibrils form, and thus help us in developing new medicines to treat
our patients with amyloidosis.
ClearVue Coverslipper
[www.thermoscientific.com/en/product/shandon-clearvue-coverslipper.html]
Jo Jerden
Dedicated funds are being raised by patient and successful fundraiser,
Joanna Jerden, for purchase of this essential piece of equipment
(www.justgiving.com/Joanna-Jerden-CoverSlipperFund). The Thermo
Scientific™ Shandon™ ClearVue™ Coverslipper is required for
histological diagnosis of amyloidosis in the National Amyloidosis
Centre. It was urgently needed before the dedicated funds became
available so the purchase was underwritten by existing funds in the
ARF, which will be fully replaced in due course so that the apparatus
will eventually have been bought solely through the efforts of those
who donated for it. Histology staff who use the automated
coverslipper in the Centre’s busy diagnostic laboratory, says that it has
greatly increased throughput and improved the accuracy of the
coverslipping workload.
FLUOstar Omega multi-mode microplate reader
[www.bmglabtech.com/products/microplate-reader/instruments.cfm?product_id=13]
This vital piece of sophisticated research equipment was purchased with
ARF money from BMG Labtech in December 2012. It is essential for a wide
range of assays performed in multiwell microtitre plates: measuring
amyloid proteins in blood and tissues in order to understand the disease
and develop new and better means of diagnosing and treating it.
Dr Riccardo Porcari
Dr Riccardo Porcari, working in Professor Vittorio Bellotti’s
Misfolding Group, is shown using this new instrument.
Protein
State-of-the-art chromatography system
[www.dionex.com/en-us/products/liquid-chromatography/lc-systems/standard/lp-72459.html]
In March 2007, ARF money was used to purchase an Ultimate 3000 system
from Dionex. This chromatography system is essential for molecular analysis
across a wide range of studies of small and large molecules involved in
amyloidosis, in order to understand the disease and develop new and better
means of diagnosis and treatment.
Dr Nigel Rendell, of the Wolfson Drug Discovery Unit, is shown using the
Ultimate system.
Dr Nigel Rendell
A very low temperature freezer
In December 2009 a -86°C upright freezer was purchased from Sanyo Gallenkamp using ARF money.
Perishable patient clinical and experimental samples can only be safely stored for testing now and in future
years by freezing them at very low temperature, for which this freezer is designed.
Electrophoresis unit
[https://ca.vwr.com/store/catalog/product.jsp?product_id=4614543]
Electrophoresis is a method for analysing and measuring a wide range of the proteins involved in amyloidosis.
In September 2009, ARF funds were used to purchase an additional Multiphor II Electrophoresis Unit plus
accessories from VWR International. This equipment is a basic essential workhorse used daily throughout the
years for these purposes.
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Sophisticated microscopy suite to test efficacy of potential new drugs
[www.mediacy.com/pdfs/IPP62Broch.pdf]
ARF funds were used to purchase an Image-Pro Plus Advanced Microscopy
Suite featuring powerful image capture and analysis tools, an QImaging Retiga
2000R high resolution digital camera and a Prior Lumen metal halide UV light
source, from Media Cybernetics Europe in June 2011. Importantly, the new
system was compatible with our existing Leica research microscope, shown
here. One of our main aims is to develop treatments which eliminate amyloid
deposits from the tissues. We therefore perform many studies testing the
efficacy of our inventions, which is measured in tissue sections by
sophisticated microscopy, for which this equipment is essential. Indeed, it
was urgently needed before dedicated funds became available so the
purchase was underwritten by the ARF. Dr Glenys Tennent, of the Wolfson
Drug Discovery Unit, who evaluated and operates the system, says that the
new system permits acquisition of consecutive image tiles to create a single
high resolution digital image montage or “virtual slice” of an entire section,
which yields more efficient and precise measurements.
STAFF AND SALARY SUPPORT
Clinical proteomics: a new diagnostic approach in amyloidosis
There are many types of proteins that form amyloid deposits, and
knowing which type is present helps us in diagnosis and determining
treatment. We currently use a number of laboratory techniques to
characterise amyloid; these include immunostaining and DNA
sequencing, as well as our highly sophisticated body scanning
techniques. We have recently introduced a new technique of
“clinical proteomics”, originally developed at the world famous
Mayo Clinic in the USA, which allows us unequivocally to identify
the proteins present in amyloid tissue.
The technique is highly sophisticated and exploits state-of-the-art
technology to the full. We use lasers to remove tiny pieces of
amyloid from tissue biopsies. Proteins within these tissue pieces are broken down into thousands of smaller
peptide fragments by enzymes, and each individual peptide is analysed and characterised using a Velos
Orbitrap mass spectrometer. Massive, complex information is then sorted, filtered and reassembled using our
dedicated bioinformatics computers, coupled with the full knowledge of the human genome, allowing us to
identify the specific proteins present in the amyloid deposits.
Velos Orbitrap mass spectrophotometer
Clinical proteomics complements and extends our current diagnostic techniques. It is fast becoming a gold
standard diagnostic test, helping us in our research into the causes of amyloidosis and, most importantly, in
providing a better and more comprehensive service for our patients. The development of clinical proteomics
at the Royal Free Hospital has been undertaken by Dr Graham Taylor and Dr Nigel Rendell, at the Wolfson
Drug Discovery Unit in a joint effort together with the NHS National Amyloidosis Centre, and demonstrates the
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the importance of applying basic scientific research to improve clinical care. Dr Taylor and Dr Rendell also use
mass spectrometry to measure the molecular mass of proteins very accurately, a critical component of many
of the other on-going research projects at the Wolfson Drug Discovery Unit.
Their ground-breaking work has been made possible by the use of the ARF to pay part of Dr Rendell’s salary
for several years. Dr Rendell is responsible for the extremely complex technical management of the mass
spectrometer, which is crucial for the success of clinical proteomics and other work with this complex
machine. His work has made possible several important discoveries and developments related both to direct
clinical diagnosis of patients and research that has led to better diagnostic methods and new and more
effective treatments. After the ARF had made Dr Rendell’s crucial work possible by paying his salary for an
initial period, core funding was obtained for the Wolfson Drug Discovery Unit from the UCLH/UCL Biomedical
Research Centre, funded by the National Institute for Health Research, and this provides his salary among
others.
Thanks to the funding from the ARF, patients at the National Amyloidosis Centre are continuing to benefit
from world class diagnostic standards which enable our doctors to provide the best possible treatment.
Amyloidosis studentship at Southampton
In 2009, a contribution from the ARF helped to support Dr Mohinder Pal, an industrious and productive
research student, enabling him to complete some research relevant to better understanding of the
pathogenesis of amyloidosis. In 2010 he completed his doctoral thesis at the University of Southampton,
entitled “Protein misfolding and human disease”. See http://eprints.soton.ac.uk/167063/.
Amyloidosis drug development collaboration between UCL and Oxford
University
The ARF contributed to the salary of Dr Julien Marcoux, a key, expert, post-doctoral scientist working on our
development of potential new drugs for amyloidosis, using unique methods developed and available only in
the laboratory of our collaborator Dame Carol Robertson FRS in Oxford.
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