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Process DEVELOPMENT
101
BULK
Group-specific Toyopearl affinity resins
Ligand Density: 7µmol/mL
N
N
NaO
3
S
NaO
3
S
NaO
3
S
NH
NH
NH
N
N
N
N
N
SO
3
Na
SO
3
Na
SO
3
Na
HW-
65 O-R-O
HO
OH
OH
N NH
N
N
Toyopearl AF-Red-650M
(1) Provided as an aqueous slurry containing 20% ethanol, v/v in 1mol/L NaCl.
(2) Provided as an aqueous slurry containing 20% ethanol.
Toyopearl AF-Blue HC-650M
HW-
65
Ligand Density: 20µmol/mL
CH
3
COO Na
CH
2
COO Na
HW-
65 O-R-O CH
2
-N
Toyopearl AF-Chelate-650M
(1)
(2)
O
O
NH
2
NH
SO
3
Na
NH
NaO
3
S
N
N
NH
SO
3
Na
N
(1)
Approximate Ligand Density: 5mg/mL
COONa
O
OSO
3
Na
CH
2
OSO
3
Na
O
HNOSO
3
Na
OH
O
OH
O
n
HW-
65
Toyopearl AF-Heparin HC-650
Activated and reactive Toyopearl affinity resins
HW-
65 O-R-O-SO
2
-CH
2
-CF
3
Ligand Density: 80 mol/g (dry)
Ligand Density: 800 mol/g (dry)
Ligand Density: 60 eq/mL
O
HW-
65 O-R-O-CH
2
-CH-CH
2
Ligand Density: 100 eq/mL
Ligand Density: 100 eq/mL
Toyopearl AF-Tresyl-650M
(1)
Toyopearl AF-Carboxy-650M
(3)
Toyopearl AF-Amino-650M
(3)
Toyopearl AF-Formyl-650M
(2)
Toyopearl AF-Epoxy-650M
(1)
(1) Provided as dry, free-flowing powder.
One gram of dry powder produces about 3.5 mL of hydrated resin.
(2) Provided as aqueous slurry, containing 1% gluteraldehyde.
(3) Provided as aqueous slurry, containing 20% ethanol.
HW-
65 O-R-O-CH
2
-CHO
HW-
65 O-R-O-CH
2
-CHOH-CH
2
NH
2
HW-
65 O-R-O-CH
2
-COOH
TOYOPEARL RESINS FOR AFC
HIGHLIGHTS
New AF-rProtein A-650F resin for antibody purification.
Active, reactive and group specific resins
Provided in standard 1000 Å pore size for high capacity of large
biopolymers.
TOYOPEARL AF-Blue HC-650M is available for albumin and interferon
applications with the lowest leaching blue.
TOYOPEARL AF-Heparin HC-650M high capacity resin exhibits an
Antithrombin III dynamic capacity of 4 mg/mL.
TOYOPEARL media for Affinity Chromatography (AFC) are based on
TOYOPEARL HW-65 resin and functionalized with either chemically
active groups or group-specific ligands. Resins with activated functional
groups are ready for direct coupling of a protein or other ligand, while
resins with reactive groups employ coupling or reductive amination
to achieve covalent bonding. The 1000 Å pore size common to all
TOYOPEARL affinity resins accommodates proteins up to 5,000,000 Da.
In general, TOYOPEARL AF-Tresyl-650M and AF-Formyl-650M are
recommended for coupling proteins, while AF-Epoxy-650M is suited
for coupling low molecular weight ligands. TOYOPEARL AF-Amino-
650M and TOYOPEARL AF-Carboxy-650M may be used in either
application. TOYOPEARL AF-Heparin HC-650M interacts with a wide
range of biomolecules including plasma components, lipoprotein lipase,
collagenase, and DNA polymerase. The structures of TOYOPEARL
activated and reactive ligands are given in
Figure 8
, while the
structures of TOYOPEARL group-specific ligands are listed in
Figure 9
.
TOYOPEARL AF-rProtein A-650F is designed for efficient and robust
purification of antibodies. The newly developed recombinant
protein A ligand is derived from one of the IgG-binding domains
of the staphylococcus aureus protein A (
Figure 10
)
.
TOYOPEARL
AF-rProtein-650F binds human and mouse immunoglobulin G with high
binding capacity and at high flow rates. This reduces column and buffer
volumes and allows fast loading procedures.
figure 9
Group-specific TOYOPEARL affinity resins
figure 10
Recombinant Protein A derived
ligand
figure 8
Activated and reactive TOYOPEARL affinity resins