Process DEVELOPMENT
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BULK
RoboColumns for toyopearl
Tosoh Bioscience offers TOYOPEARL media now also in the well-
known RoboColumn® format packed by Atoll GmbH. RoboColumns
are miniaturized chromatographic columns pre-packed with the
most popular TOYOPEARL ion exchange, mixed-mode, hydrophobic
interaction or affinity media. The columns are available in different
volumes and can be operated with a robotic liquid handling system.
This approach allows automated high-throughput, small-scale
chromatographic separations of protein samples by running up to eight
individual columns simultaneously.
RoboColumns are available in two formats with 200 μL (bed height of
10 mm) and 600 μL (bed height of 30 mm) resin volume, respectively.
They are supplied in a row of eight units pre-packed with the same
TOYOPEARL resin and sealed with two removable silicon cover seals
for proper storage. A 96-well array plate is available to arrange the up
to 96 RoboColumn units.
Figure 1 shows a screening experiment to optimize the parameters
for the intermediate flow-through anion exchange step in a mAb
purification platform. Protein binding of a Protein A capture eluate on
RoboColumns packed with TOYOPEARL SuperQ-650M was analyzed by
varying salt concentration and pH of loading and washing buffer. Best
results were achieved using 20 mmol/L sodium phosphate, 80 mmol/L
sodium chloride, pH 6.5.
figure 5
Optimization of anion exchange conditions
0
100
200
300
400
500
600
10
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1
10mM PBS/40mM NaCI/pH 7.5
10mM PBS/40mM NaCI/pH 7.5
20mM PBS/80mM NaCI/pH 7.5
30mM PBS/120mM NaCI/pH 7.5
10mM PBS/10mM NaCI/pH 6.5
10mM PBS/40mM NaCI/pH 6.5
20mM PBS/80mM NaCI/pH 6.5
30mM PBS/120mM NaCI/pH 6.5
V
[µl]
STRIP
PLW
LOAD
C (mg/mL)
Elution profile of a protein A capture eluate on RoboColumns packed with
Toyopearl SuperQ-650M at various conditions. Data kindly provided by
T. Schröder, Atoll GmbH.
FEATURES
BENEFITS
Pre-packed columns for use with robotic systems
High throughput parallel chromatography
200 and 600 µl bed volume
Fast resin screening and evaluation of design space
Miniaturized column format
Low consumption of sample and buffers
