65
afc
Affinity
chromatography
TOYOPEARL RESINS FOR AFFINITY LIGAND COUPLING
TOYOPEARL offers a spectrum of carefully selected affinity
resins primed with activated or reactive groups which can
be used to covalently attach almost any custom ligand. The
structures of TOYOPEARL activated and reactive ligands
are shown in Figure 1.
In general, TOYOPEARL AF-Tresyl-650M and TOYOPEARL
AF-Formyl-650M resin are recommended for coupling
proteins, while TOYOPEARL AF-Epoxy-650M resin is suited
for coupling lower molecular weight ligands. TOYOPEARL
AF-Amino-650M and TOYOPEARL AF-Carboxy-650M resins
may be used for both.
TOYOPEARL affinity resins may be used in combinato-
rial chemistry or
for solid phase synthesis of peptides and
oligonucleotides because of their excellent stability in a
variety of organic solvents and under extremes of pH.
figure 1
ACTIVATED TOYOPEARL AFFINITY RESINS
Toyopearl AF-Tresyl-650M
Toyopearl AF-Epoxy-650M
HW-
65 O-R-O-SO
2
-CH
2
-CF
3
Ligand Density: 80 µmol/g (dry)
Ligand Density: 800 µmol/g (dry)
Toyopearl AF-Formyl-650M
Ligand Density: 60meq/L
O
HW-
65 O-R-O-CH
2
-CH-CH
2
HW-
65 O-R-O-CH
2
-CHO
Toyopearl AF-Amino-650M
Ligand Density: 100mmol/L
HW-
65 O-R-O-CH
2
-CH-CH
2
-NH
2
OH
Toyopearl AF-Carboxy-650M
HW-
65 O-R-O-CH
2
COOH
Ligand Density: 100 meq/L
REACTIVE TOYOPEARL AFFINITY RESINS
Table II
Representative coupling densities for activated and reactive TOYOPEARL media
TOYOPEARL resin
AF-Tresyl-650M
AF-Formyl-650M
AF-Amino-650M
AF-Carboxy-650M
Protein coupled (mg/mL resin)
Soybean trypsin inhibitor
16
3.5
5.8
15
Protein A
1.9
-
-
-
Concanavalin A
13
-
-
-
α
1-Antitrypsin
12.3
-
-
-
α
-Chymotrypsin
12.5
-
-
-
Myoglobin
12.4
-
-
-
Ovalbumin
-
2.5
6.7
0.8
Bovine serum albumin
12.4
14
19.2
3.3
Human IgG
10.0
15
6.7
11.7
Cytochrome C
-
5.8
3.3
7.5
Lysozyme
60
20
5.8
17.5
Coupling agent
not required
NaCNBH
3
NaCNBH
3
or Carbodiiamide
Carbodiimide
Optimal pH
7.0 - 9.0
6.9 - 9.0
4.5 - 6.0
4.5 - 6.0
