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BULK
TOYOSCREEN PROCESS DEVELOPMENT COLUMNS
ToyoScreenProcessDevelopment columnsareeasy-to-use, pre-packed
columns containing Tosoh Bioscience‘s most popular TOYOPEARL
resins. These columns provide a convenient, low-cost method for the
evaluation of TOYOPEARL ligand chemistries. ToyoScreen Process
Development columns are available in packages of 6 x 1 mL and 6 x
5 mL volumes for affinity, ion exchange and hydrophobic interaction
chromatography. For the new TOYOscreen AF-rProtein A-650F
package sizes are 5 x 1 mL, 1 x 5 mL and 5 x 5 mL. See the chapter on
bulk resins for detailed information on TOYOPEARL resins.
Screening
Historically, resin screening was accomplished by manually packing
various bulk resins into small columns requiring a significant investment
in time and cost. In order to improve the efficiency of resin screening
experiments, pre-packed ToyoScreen Process Development columns
were developed for the evaluation of different TOYOPEARL resins.
Scalability
Initial results from resin screening and optimization with ToyoScreen
columns can accurately predict the separation behavior at larger
scales.
Figure 1
illustrates the similar retention time behavior between
1 mL ToyoScreen columns and con-ventional 7.5 mm ID x 7.5 cm L
analytical columns. Additionally,
Figure 2
depicts a practical antibody
scale up in which conditions were set using a 1 mL ToyoScreen column
and applied to a 10 mL semi-preparative column with a different inner
diameter and length. Similar resolution results are predicted by the
following equation:
Method Optimization
Besides the determination of what sticks during resin screening
experiments, ToyoScreen Process Development columns can be used
to quickly establish optimum elution conditions. Varying pH, salt type,
salt gradients and flow rate are common experimental parameters
explored. The effect of varying salt type and pH are shown in
Figures
3 &
4
for anti-TSH in cell culture supernatant on ToyoScreen Phenyl-
650M.
figure 1
Comparison of selectivity between Toyoscreen and Conventional
Column
0
10
20
30
40
50
Ether-650M
PPG-600M
Phenyl-650M
Butyl-650M
SuperButyl-550C
Hexyl-650C
Retention Time (min)
Conventional (
α
-ChymotrypsinogenA)
ToyoScreen (
α
-ChymotrypsinogenA)
Conventional (Lysozyme)
ToyoScreen (Lysozyme)
Conventional (RibonucleaseA)
ToyoScreen (RibonucleaseA)
Columns: ToyoScreen (1 mL), Conventional Column (7.5 mm ID x 7.5 cm L)
Eluent A: 0.1 mol/L phosphate buer + 1.8 mol/L sodium sulfate (pH 7.0)
Eluent B: 0.1 mol/L phosphate buer (pH 7.0)
Flow Rate: 1 mL/min
Gradient: 30 min linear
Inj. Vol.: 50 µL
Samples: Ribonuclease A, Lysozyme,
α
-Chymotrypsinogen, 1 mg/mL
Retention time of conventional column was plotted after converting
following equation: plotted value = actual measurement value - 4.82
Comparison of selectivity between ToyoScreen
and Conventional Column
FEATURES
BENEFITS
Pre-packed columns
Easy to set up and screen an entire resin series for a specific
chromatographic mode
1 mL and 5 mL bed volume
For sample limited applications with up to milligram purifications
Cartridge design
Provides low cost, efficient alternative to hand packing with bulk
resin
Ready to connect to ÄKTA, FPLC and HPLC systems
Seamless integration into any platform
Six pieces offered in mixed or single chemistry
For cost savings in screening or process experiments
Columns: ToyoScreen (6.4 mm ID x 3 cm L), Conventional Column (7.5 mm
ID x 7.5 cm L);
Eluent A: 0.1 mol/L phosphate buffer + 1.8 mol/L sodium sulfate (pH 7.0),
Eluent B: 0.1 mol/L phosphate buffer (pH 7.0); Flow Rate: 1 mL/min
Gradient: 30 min linear; Inj. Vol.: 50 μL; Samples: Ribonuclease A, Lysozyme,
a
-Chymotrypsin gen, 1 mg/mL
Retention time of conventional column was plotted after converting following
equation: plotted value = actual measurement value - 4.82