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Process DEVELOPMENT
99
BULK
TOYOPEARL AND TSKgel BULK RESINS FOR IEC
HIGHLIGHTS
TOYOPEARL GigaCap
®
S-650M, CM-650M and Q-650M resins are
high capacity ion exchange resins featuring high dynamic binding
capacities for both small molecules like insulin and larger proteins
like monoclonal antibodies.
Weak and strong anion and cation exchangers are offered in both
product lines.
Standard 1,000 Å pore size for large biopolymers and 500 Å pore size
packing for optimal binding capacity are available.
High efficiency TSKgel resins scale up directly from TSKgel analytical
columns.
For separating mixtures of biomolecules, Ion Exchange Chromatography
(IEC) is known for its high resolution and high capacity. It is very
effective in the initial capture step of a chromatography process. IEC is
also useful for further purification and/or polishing. It can complement
other chromatographic techniques in the design of an economical
downstream purification process. IEC is often used as a purification
step before HIC, SEC, and RPC. IEC will also purify and concentrate the
target molecule in one step when the sample is diluted. This also allows
it to be used as a concentration step after SEC.
A 5000-fold scale-up of a
α
-galactosidase enzyme purification was
accomplished using TOYOPEARL DEAE-650M. The chromatograms
in
Figure 6
demonstrate the excellent scale up characteristics of
TOYOPEARL ion exchange media. Gradient slope and particle diameter
remained unchanged. Linear velocity was reduced by 15% in the largest
scale separation, and resolution actually improved relative to the
smallest scale separation. This may be partly attributed to increased
bed height and the slower linear velocity. Although the column volume
was increased in part by increasing the bed height, the principal change
in column volume was a result of the greater column diameter (1.4 cm
to 60 cm L). This example illustrates how TOYOPEARL media can be
conveniently scaled up from laboratory to production scale applications
using the same particle size if desired.
Because the correct choice of an ion exchange resin can have
a considerable impact on the economy of a process, Tosoh Bioscience
provides many product options in both TOYOPEARL and TSKgel IEC
bulk polymeric media. See
Table III
for a complete listing of available
particle sizes. Ordering information for quantities < 1 L is provided at the
end of this section.
figure 6
Process scale-up purification of
b
-galactosidase with TOYOPEARL
DEAE-650M
Process scale-up purification of
b
-
galactosidase with
Toyopearl DEAE-650M
Column:
Sample:
Elution:
Flow rate:
Linear velocity:
Detection:
Toyopearl DEAE-650M
1%
b
-galactosidase: A. 8mL; B. 1L; C. 40L
linear gradient from 0.03 to 0.10mol/L NaCl
in 0.014mol/L Tris-HCl (pH7.7)
A. 1.0mL/min; B. 60mL/min; C. 1.6L/min
A. 39cm/h; B. 40cm/h; C. 34cm/h
UV @ 254nm
B. 2.7 L column
(108 mm ID x 30 cm)
A. 23mL column
(14 mm ID x 15 cm)
C. 113 L column
(600 mm ID x 40 cm)
Relative Elution Volume (Ve /V v )
Absorbance at 254 nm
0
5
10
15
Description
Type*
Part. size (µm)
Anion Exchange
TSKgel DEAE-5PW
W
20, 30
TSKgel SuperQ-5PW
S
20, 30
TOYOPEARL DEAE-650
W
35, 65, 100
TOYOPEARL SuperQ-650
S
35, 65, 100
TOYOPEARL QAE-550
S
100
TOYOPEARL Q-600 AR
S
100
TOYOPEARL GigaCap Q-650M
S
75
Cation Exchange
TSKgel SP-5PW
S
20, 30
Tskgel SP-3PW
S
30
TOYOPEARL CM-650
W
35, 65, 100
TOYOPEARL GigaCap CM-650M
W
75
TOYOPEARL SP-550
S
100
TOYOPEARL SP-650
S
35, 65, 100
TOYOPEARL MegaCap II SP-550EC
S
100-300
TOYOPEARL GigaCap S-650M
S
75
*W = Weak; S = Strong
TABLe III
TOYOPEARL and TSKgel Ion Exchange Resins
Column: TOYOPEARL DEAE-650 M; Sample: 1%
b
-galactosidase: A. 8 mL; B.
1L; C. 40L Elution: linear gradient from 0.03 to 0.10 mol/L NaCl in 0.014 mol/L
Tris-HCl (pH7.7); Flow rate: A. 1.0 mL/min; B. 60 mL/min; C. 1.6 L/min; Linear
velocity: A. 39 cm/h; B. 40 cm/h; C. 34 cm/h; Detection: UV@254nm