68
afc
Affinity
chromatography
figure 7
group-specific toyopearl affinity resin
GROUP-SPECIFIC TOYOPEARL AFFINITY RESINS
Approximate Ligand Density: 5 g/L
Toyopearl AF-Heparin
COONa
O
OSO
3
Na
CH
2
OSO
3
Na
O
HNOSO
3
Na
OH
O
OH
O
n
HW-
65 O-R-O
Ligand Density: 20 mmol/L
CH
2
COO Na
CH
2
COO Na
HW-
65 O-R-O CH
2
-N
Toyopearl AF-Chelate
N
N
N
R
2
R
2
NH
O-R-O
HW-
65
NH
NaO
3
S
SO
3
Na
NH
2
NH O
O
Toyopearl AF-Blue HC
R
1
=H or SO
3
Na
R
2
=H or SO
3
Na
Ligand Density: 15 mmol/L
Toyopearl AF-Red
NaO
3
S
SO
3
Na
N
N
N
N
NaO
3
S
NaO
3
S
OH
N
N
N
NH
HO
N
N
N
NH
NH
SO
3
Na
SO
3
Na
NH
OH
HW-
65
TOYOPEARL resins with group specific ligands
The structures of TOYOPEARL group specific ligands are
shown in Figure 7.
TOYOPEARL AF-Chelate-650M
This resin is derivatized with iminodiacetic acid (IDA) at a
concentration of ca. 20 µmol/mL. In typical applications,
selected metal ions, most often Ca
2+
, Ni
2+
, Zn
2+
, Co
2+
and Cu
2+
are bound to the support by stable chelation. The resultant
metal ion-bearing resin binds to histidine and free cysteine
containing sequences of a peptide or protein. Immobilized
metal ion affinity chromatography (IMAC) has been used
for purification of recombinant human growth factor, tissue
plasminogen activator, glycophorins, and whole cells.
TOYOPEARL AF-Blue HC-650M
Functionalized with Cibachron Blue F3G-A, TOYOPEARL
AF-Blue HC-650M resin has excellent capacity for proteins,
particularly albumin (Figure 8). In addition, this high
capacity resin has improved caustic stability, reduced dye
ligand leakage, and superior pressure-flow characteristics
relative to more traditional agarose materials (Figure 9).
TOYOPEARL AF-Red-650ML
Toyoscreen and TOYOPEARL AF-Red-650ML resins are
functionalized with Procion Red HE-3B, (also known as
Reactive Red 120). This resin is useful for the purification of
nucleotide dependent enzymes, lipoproteins, plasminogen,
peptides, hormones and cytotoxins.
These two dye-ligand resins are useful in binding/purifica-
tion of nucleotide-dependent enzymes, albumin, cell growth
factors, interferons, transferases, cyclases. and polymer-
ases. Typical binding capacities are shown in Table III.
TOYOPEARL AF-Heparin HC-650M
Heparin is a linear and highly sulfated glycosaminoglycan
which has anti-coagulant properties. Due to its polyanionic
nature, heparin interacts with a wide range of biomolecules
including plasma components, lipoprotein lipase, collage-
nase, and DNA polymerase.
Immobilized heparin is widely used as an adsorbent in
affinity chromatography for the purification of biological
substances. TOYOPEARL AF-Heparin HC-650M resin is a
high capacity, affinity
adsorbent with excellent chemical
stability.
