USD2992(4)
ScreenExpert RoboColumn◆
For High Throughput Chromatography Sorbent
Screening and Optimization
ScreenExpert RoboColumns are miniaturized
chromatography columns pre-packed with Pall
chromatography sorbents. They are designed
for fully automated and parallel chromatographic
separations with robotic liquid handling workstations, such as the Freedom EVO◆ from Tecan.
The 200 µL column volume is recommended
for sorbent and process conditions screening,
while the 600 µL is used for purification process
optimization or when a residence time above
4 minutes is required (e.g., capacity study on
mixed-mode sorbents).
ScreenExpert RoboColumns◆ are packed with
Pall anion exchange, mixed-mode, affinity or any
other Pall chromatography sorbents upon request.
The column units are packed by Atoll GmbH and
are identical to Atoll’s MediaScout◆ RoboColumns.
The columns are supplied as rows of eight
pre-packed units, with two removable silicon
cover seals for proper storage. A 96-well array
plate is available to arrange up to 96 ScreenExpert
RoboColumn units.
Two formats are available, 200 µL and 600 µL,
with a bed height of 10 and 30 mm respectively.
Refer to Table 1 for characteristics and available
column formats.
Benefits
Applications
Column format, and high quality packing enables the ability
to mimic all the steps of a chromatography separation in
dynamic mode. Data obtained are comparable to those
obtained in larger columns
ScreenExpert RoboColumns are suitable for a large variety
of applications such as:
Miniaturized format facilitates screening of chromatography
sorbents, reduces sample consumption, and saves time
Parallel screening and optimization of chromatographic
conditions in downstream process development
(conductivity, pH)
Perfectly fitted to Design of Experiment (DoE) requirements,
allowing a large amount of experimental work in a minimum
of time
96-well format allows easy integration with automation
systems for High Throughput Process Development (HTPD)
Multiple parameters can be tested in parallel, e.g.,
residence time
Reusable
Flexible use: the user makes column selection when needed
Table 1
ScreenExpert RoboColumns Characteristics
ScreenExpert RoboColumns
200 µL
600 µL
10 mm
30 mm
5 mm
Polypropylene
All commonly used aqueous buffers, pH 1 to 14
Halogenated organic solvents, hexane
20% ethanol /1 M NaCl
2 to 8 °C
4 to 30 °C
Up to 8.0 bar g (116 psi g)
16 to 1,000 cm/hr(1)
Placed on every individual column body, and
containing:
• Product number
• Sorbent name
• Column volume in µL
96-Well Array Plate
Size
Material
Number of wells
128.3 x 86.0 x 14.0 mm
Polyoxymethylene copolymer
96
(1) The Liquid Transfer menu of the Te-Chrom◆ Wizard enables set up of
the flow rate for each individual dispensing and chromatography step
(5.1.1.6 Liquid Transfer, Te-Chrom Wizard Software Manual, 396076,
en, V1.0).
2
Scale down experimental work
Determination of dynamic binding capacity
Determination of the optimal residence time
Optimization of the regeneration step
Application Example: Development of the Capture Step of
a Monoclonal Antibody (MAb) Expressed in a Crude CHO
Feedstock
MAb binding conditions were tested on 3 mixed-mode sorbents
and dynamic binding capacity (DBC) was evaluated based on
breakthrough (BT) curve (Figure 1).
Characteristics
Column volume
Bed height
Column inner diameter
Column material
Chemical stability
Avoid
Storage solution
Recommended storage
temperature
Working temperature
Maximum working pressure
Flow rate
Laser label
Parallel screening of chromatography sorbents
The ScreenExpert RoboColumn was overloaded with MAb CHO
feedstock at 0.52 mg/mL and flowthrough (FT) was collected
as one-column-volume (CV) fractions (200 µL each) in 350 µL
96-well plate. The amount of MAb recovered in the FT fractions
was evaluated using Bio-Layer Interferometry (BLI) with protein A
sensor (Octet® System, Pall ForteBio). It was plotted against the
volume of feedstock loaded per ScreenExpert RoboColumn unit.
1/10 of the MAb concentration (C/10) was 0.052 mg/mL
(52 µg/mL in Figure 1). The volume equivalent at 10% BT
(Veq10%BT) was the volume of MAb loaded when reaching
C/10 in FT fractions (8.56 mL in Figure 1).
Figure 3
DBC on MEP HyperCel Sorbent in ScreenExpert RoboColumn
and in PRC Column
Figure 1
Breakthrough Curve on MEP HyperCel Sorbent in 200 µL
ScreenExpert RoboColumn
25
DBC 10% BT (mg MAb/mL Sorbent)
MAb Concentraton in Flowthrough
µg/mL
130
110
90
70
C/10 50
30
10
0
-10
-30
0
2
4
6
8
10
Veq 10% BT
MAb CHO Feedstock Volume Loaded
15
10
5
0
ScreenExpert RoboColumn
PRC Column
A 335-fold scale-up of the MAb capture step was finally carried
out from 200 µL ScreenExpert RoboColumn to a lab-scale
column (2.5 cm I.D) packed with 67 mL MEP HyperCel sorbent
(Figure 4).
DBC10% BT (mg/mL sorbent) =
Veq10%BT (mL)* x MAb concentration (mg/mL)
Column volume (mL)
*void volume was taken into account
Using this breakthrough curve method, the DBC for MEP, HEA
and PPA HyperCel sorbents was determined on 200 µL
ScreenExpert RoboColumns (Figure 2).
Figure 2
DBC on 3 Mixed-Mode HyperCel Sorbents in ScreenExpert
RoboColumns for Different Binding Conditions
This comparison showed that MAb binding was maximized on
DBC10% BT (mg MAb/mL Sorbent)
20.9
20
mL
Breakthrough curve during the loading of a MAb CHO feedstock
(0.52 mg/mL) at a ~4-minute residence time.
25
21.9
21.9
Figure 4
335-fold Scale-Up of the MAb Capture Step from ScreenExpert
RoboColumn to a 2.5 cm ID Lab-Scale Column
FT
mAU
W
E1
E2
E3
pH
4000
Abs. at 280 nm
pH
Conductivity
3500
3000
10.0
8.0
2500
2000
6.0
1500
20
1000
15
14.6
13.0
10.0
10
10.0
4.0
500
0
0
500
1000
1500
2000
2500
3000
5
0
MEP HyperCel
PBS, pH 7.4
MEP HyperCel
1 M NaCl, pH 7.4
HEA HyperCel
1 M NaCl, pH 7.4
HEA HyperCel
1 M NaCl, pH 8.5
PPA HyperCel
PBS, pH 7.4
MEP HyperCel sorbent in PBS, pH 7.4. The selected condition
was then transferred to 1 mL PRC column for confirmation of
the DBC, approximately 21 mg/mL (Figure 3).
mL
Chromatogram of MAb Capture Step on a 2.5 cm ID lab-scale column
(67 mL). Equilibration in PBS, pH 7.4. Loading of MAb feedstock =
21 mg MAb/mL MEP HyperCel sorbent. Residence time = 4 minutes;
FT: Flowthrough; W: Wash; E1, E2 and E3: Elution at pH 5, 4 and 3
respectively.
The analysis of fractions obtained on ScreenExpert RoboColumn
and a 2.5 cm ID lab-scale column were in the same magnitude.
Comparable data in terms of recovery (100%) and purity (90%)
were evidenced (Figure 5, Table 2).
www.pall.com/biopharm
3
Figure 5
Fraction Analysis From ScreenExpert RoboColumn and a 2.5 cm
ID Lab-Scale Column Using BLI
ScreenExpert RoboColumn
nm
7
E1
6
F0
5
4
ScreenExpert
RoboColumn
Lab Scale
Column
2.5 cm I.D.
Purity
(%)**
0.2
4
22
102
86
66.7
1376
21
99
91
* Evaluated using BLI technology (Octet System, Pall ForteBio)
** Evaluated using LabChip◆ GXII technology (PerkinElmer)
3
E2
2
1
E3
W
FT
0
-1
0
20
40
60
Time (sec)
80
100
120
2.5 cm ID Lab-Scale Column
nm
7
E1
F0
6
5
4
3
2
E2
W
FT
E3
1
0
-1
0
20
40
60
Time (sec)
80
100
120
Interaction of the MAb with protein A sensor was measured using BLI.
Signal is correlated to MAb concentration using a standard curve
(data not shown). Residence time = 4 minutes. F0: Sample load;
FT: Flowthrough; W: Wash; E1, E2 and E3: Elution at pH 5, 4 and
3 respectively.
4
Table 2
Summary of the MAb Capture Step on ScreenExpert
RoboColumn or a 2.5 cm ID Lab-Scale Column
MAb
Loading
Column
Volume Loaded
(mg MAb/
Recovery
Type
(mL)
(mg)*
mL sorbent) (%)*
Handling
ScreenExpert RoboColumns are designed for robotic handling.
When using the high-throughput process development approach
for ScreenExpert RoboColumns, it is suggested to use Design
of Experiments (DoE) criteria for the experimental set-up. This
enables to screen many different chromatographic parameters
such as resin type, pH, conductivity / ionic strength, etc.,
efficiently and simultaneously.
For experimental set-up and the configuration of the 96-well
array plate, the Te-Chrom Wizard is recommended. The
Te-Chrom Wizard is a dialog-based graphic user interface
offered by Tecan and based on the Freedom EVOware
software. The Te-Chrom Wizard was developed for the use
of ScreenExpert RoboColumns and provides the possibility
to configure hardware, plate layout, and the chromatographic
process as well as to set all process relevant chromatographic
parameters, e.g., flow rate, volume without direct script writing.
Ordering Information
Complementary Products
Part
Number
Description
ScreenExpert RoboColumn MEP HyperCel 200 µL, row of 8
ScreenExpert RoboColumn MEP HyperCel 600 µL, row of 8
ScreenExpert RoboColumn HEA HyperCel 200 µL, row of 8
ScreenExpert RoboColumn HEA HyperCel 600 µL, row of 8
ScreenExpert RoboColumn PPA HyperCel 200 µL, row of 8
ScreenExpert RoboColumn PPA HyperCel 600 µL, row of 8
ScreenExpert RoboColumn HyperCel STAR AX 200 µL,
row of 8
ScreenExpert RoboColumn HyperCel STAR AX 600 µL,
row of 8
ScreenExpert RoboColumn CM Ceramic HyperD F 200 µL,
row of 8
ScreenExpert RoboColumn CM Ceramic HyperD F 600 µL,
row of 8
ScreenExpert RoboColumn HyperCel STAR CEX 200 µL,
row of 8
ScreenExpert RoboColumn HyperCel STAR CEX 600 µL,
row of 8
ScreenExpert RoboColumn CMM HyperCel 200 µL, row of 8
ScreenExpert RoboColumn CMM HyperCel 600 µL, row of 8
ScreenExpert RoboColumn KANEKA KanCapA 200 μL,
row of 8
ScreenExpert RoboColumn KANEKA KanCapA 600 μL,
row of 8
96-well RoboColumn array plate
SR2MEP
SR6MEP
SR2HEA
SR6HEA
SR2PPA
SR6PPA
SR2STARAX
AcroPrep ScreenExpert Plates
Description
Part Number
MEP, HEA and PPA HyperCel Mixed-Mode Sorbents 96WP3MIXM50
MEP HyperCel Mixed-Mode Sorbent
96WPMEP50
HyperCel STAR AX Anion Exchange Sorbent
96WPSTARAX50
Multi-Well Plates (empty)
AcroPrep Advance 96-Well Filter Plate, 1 mL,
0.45 μm Supor® membrane (5/pkg)
8129
SR6STARAX
SR2CMCHDF
SR6CMCHDF
SR2STARCEX
AcroPrep Advance 96-Well Filter Plate for Ultrafiltration
Description
Part Number
350 μL, Omega™ 3K MWCO (10/pkg)
350 μL, Omega 10K MWCO (10/pkg)
350 μL, Omega 30K MWCO (10/pkg)
350 μL, Omega 100K MWCO (10/pkg)
8033
8034
8035
8036
SR6STARCEX
SR2CMM
SR6CMM
SR2KANCAPA
SR6KANCAPA
SR96WAP
PRC Prepacked Columns
Description
Part Number
PRC Column 5x50 MEP HyperCel, 1 mL
PRC Column 8x100 MEP HyperCel, 5 mL
PRC Column 5x50 HEA HyperCel, 1 mL
PRC Column 8x100 HEA HyperCel, 5 mL
PRC Column 5x50 PPA HyperCel, 1 mL
PRC Column 8x100 PPA HyperCel, 5 mL
PRC Column 5x50 HyperCel STAR AX, 1 mL
PRC Column 8x100 HyperCel STAR AX, 5 mL
PRC Column 5x50 Q Ceramic HyperD F, 1 mL
PRC Column 5x50 CM Ceramic HyperD F, 1 mL
PRC Column 5x50 HyperCel STAR CEX, 1 mL
PRC Column 8x100 HyperCel STAR CEX, 5 mL
PRC Column 5x50 CMM HyperCel, 1 mL
PRC Column 8x100 CMM HyperCel, 5 mL
PRC Column 5x50 KANEKA KanCapA, 1 mL
PRC Column 8x100 KANEKA KanCapA, 5 mL
PRC05X050MEPHCEL01
PRC08X100MEPHCEL01
PRC05X050HEAHCEL01
PRC08X100HEAHCEL01
PRC05X050PPAHCEL01
PRC08X100PPAHCEL01
PRCSTARAX1ML
PRCSTARAX5ML
PRC05X050QCHDF01
PRC05X050CMCHDF01
PRCSTARCEX1ML
PRCSTARCEX5ML
PRCCMMHCEL1ML
PRCCMMHCEL5ML
PRC05X050KANCAPA
PRC08X100KANCAPA
Visit us on the Web at www.pall.com/biopharm
E-mail us at biopharm@pall.com
Corporate Headquarters
Port Washington, NY, USA
+1.800.717.7255 toll free (USA)
+1.516.484.5400 phone
biopharm@pall.com e-mail
European Headquarters
Fribourg, Switzerland
+41 (0)26 350 53 00 phone
LifeSciences.EU@pall.com e-mail
Asia-Pacific Headquarters
Singapore
+65 6389 6500 phone
sgcustomerservice@pall.com e-mail
International Offices
Pall Corporation has offices and plants throughout the world in: Argentina, Australia,
Austria, Belgium, Brazil, Canada, China, France, Germany, India, Indonesia, Ireland, Italy,
Japan, Korea, Malaysia, New Zealand, Norway, Philippines, Poland, Russia, Singapore,
South Africa, Spain, Sweden, Switzerland, Taiwan, Thailand, United Kingdom, and
Vietnam. Distributors in all major industrial areas of the world. To locate the Pall office
or distributor nearest you, visit www.pall.com/contact.
The information provided in this literature was reviewed for accuracy at the time
of publication. Product data may be subject to change without notice. For current
information consult your local Pall distributor or contact Pall directly.
© 2016, Pall Corporation. Pall,
, HyperD, HyperCel, Octet, Omega and Supor are trademarks of
Pall Corporation. ® indicates a trademark registered in the USA and TM indicates a common law trademark.
Filtration. Separation. Solution. is a service mark of Pall Corporation. ◆MediaScout, RoboColumn and
RoboColumns are trademarks of Atoll GmbH. LabChip is a trademark of PerkinElmer. Freedom EVO,
Freedom EVOware, and Te-Chrom are trademarks of Tecan Group. KANEKA KanCapA is a trademark of
KANEKA Corporation.
1/16, PDF, GN16.9894
USD2992(4)