Filler for affinity chromatography
Provided is a filler for affinity chromatography having a high dynamic binding capacity for proteins and having excellent alkali resistance and storage stability. The filler for affinity chromatography of the present invention comprises a porous particle consisting of a copolymer of 40 parts to 99.5 parts by mass of (M-1) a methacryloyl group-containing vinyl monomer that contains a hydroxyl group and does not contain an epoxy group, 0.5 parts to 30 parts by mass of (M-2) an epoxy group-containing vinyl monomer, 0 parts to 59.5 parts by mass of (M-3) a methacryloyl group-containing vinyl monomer which is other than the monomers (M-1) and (M-2), and 0 parts to 25 parts by mass of (M-4) a vinyl monomer other than the monomers (M-1), (M-2) and (M-3) (with the proviso that the total amount of the contents of (M-1), (M-2), (M-3) and (M-4) is 100 parts by mass); ring-opened epoxy group obtainable by ring-opening of epoxy group that is contained in the copolymer; and ligand that is bound to the porous particle.
1. A filler, comprising:
a porous particle comprising a copolymer comprising:
(a) 40 to 99.5 parts by mass of (M-1) a methacryloyl group-comprising vinyl monomer comprising a hydroxyl group and not comprising an epoxy group;
(b) 0.5 to 30 parts by mass of (M-2) an epoxy group comprising vinyl monomer;
(c) 0 to 59.5 parts by mass of (M-3) a methacryloyl group-comprising vinyl monomer which is different from the monomers (M-1) and (M-2); and
(d) 0 to 25 parts by mass of (M-4) a vinyl monomer different from the monomers (M-1), (M-2), and (M-3),
with the proviso that a total amount of (M-1), (M-2), (M-3), and (M-4) is 100 parts by mass;
a ring-opened epoxy group obtained by ring-opening an epoxy group contained in the copolymer using a blocking agent having a mercapto group; and
a ligand, which is bound to the porous particle.
2. The filler of claim 1 , wherein the ligand is an alkali resistant immunoglobulin binding protein.
3. The filler of claim 1 , wherein the monomer (M-1) has formula (A):
H 2 C═C(CH 3 )C(═O)OCH 2 CH(OH)—R (A),
wherein R is a hydrogen atom or a monovalent organic group.
4. The filler of claim 3 , wherein the formula (A), R is a hydroxymethyl group or a methacryloyloxymethyl group.
5. The filler of claim 1 , wherein the particle size of the porous particle is in a range from 35 μm to 100 μm.
6. A method for isolating an immunoglobulin, the method comprising:
performing affinity chromatography with the filler of claim 1 , to adsorb an immunoglobulin to the filler;
eluting the immunoglobulin; and
washing the filler with an alkaline liquid.
7. A packed column, which is packed with the filler of claim 1 .
8. The filler of claim 3 , wherein, in formula (A), R is a hydroxymethyl group.
9. The filler of claim 3 , wherein, in formula (A), R is a methacryloyloxymethyl group.
10. The filler of claim 1 , wherein the content of the monomer (M-2) in the copolymer of the porous particle is from 1 to 20 parts by mass.
11. The filler of claim 1 , wherein the monomer (M-3) is present in the copolymer of the porous particle, and present in an amount up to 50 parts by mass.
12. The filler of claim 1 , wherein the content of the monomer (M-1) in the copolymer of the porous particle is from 45 to 96 parts by mass.
13. The filler of claim 1 , wherein the monomer (M-4) is present in the copolymer of the porous particle, and present in an amount up to 25 parts by mass.
14. The filler of claim 11 , wherein the monomer (M-4) is present in the copolymer of the porous particle, and present in an amount up to 25 parts by mass.
15. The filler of claim 1 , wherein the particle size of the porous particle is in a range from 38 μm to 75 μm.
16. The filler of claim 1 , wherein ring-opened epoxy group is ring-opened by using mercaptoethanol or thyoglycerol.