Process DEVELOPMENT
115
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-HC 650F resin for antibody purification.
Active, reactive and group specific resins
Provided in standard 100 nm 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 100 nm 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 10
, while the
structures of TOYOPEARL group-specific ligands are listed in
Figure 11
.
TOYOPEARL AF-rProtein A HC-650F designed for efficient and
robust purification of antibodies. The newly developed recombinant
protein A ligands are derived from one of the IgG-binding domains
of the staphylococcus aureus protein A (
Figure 12
)
.
TOYOPEARL
AF-rProtein A HC-650F binds immunoglobulin G with high binding
capacity and at high flow rates. This reduces column and buffer volumes
and allows fast loading procedures.
figure 11
Group-specific TOYOPEARL affinity resins
figure 12
Recombinant Protein A derived ligand
figure10
Activated and reactive TOYOPEARL affinity resins
