Abstract: There are provided a solid-phase carrier and a carrier for affinity refining that exhibit high dynamic binding capacity when a ligand is immobilized, have excellent antifouling properties, and are unlikely to have non-specific adsorption of impurities. The solid-phase carrier has a functional group capable of fixing a ligand and is characterized by having a group having a sulfinyl group, a sulfide group, an oxy group or an imino group, and a hydroxyl group, a thiol group, an amino group, a carboxy group, a sulfate group, a phosphate group or an alkanoyl group.

Abstract: 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.

Abstract: Provided is a filler for affinity chromatography which is useful for protein purification and contains porous particles that have a high dynamic binding capacity for proteins and excellent pressure characteristics. The filler for affinity chromatography of the present invention is characterized in that it includes a porous particle consisting of a polymer of vinyl monomer including a cross-linkable vinyl monomer that contains hydroxyl group but does not contain epoxy group and an epoxy group-containing non-cross-linkable vinyl monomer, or a cross-linkable vinyl monomer that contains hydroxyl group and epoxy group, ligands bound to the porous particle, and ring-opened epoxy groups.

Abstract: Provided are a filler for affinity chromatography which has excellent alkali resistance, and a method for isolating immunoglobulin. The filler for affinity chromatography is a filler in which a protein represented by the following formula (1) is immobilized on a carrier. R—R2??(1) wherein R represents an amino acid sequence consisting of 4 to 300 amino acid residues containing a region consisting of 4 to 20 contiguous histidine residues; and R2 represents an amino acid sequence capable of binding to immunoglobulin, the amino acid sequence consisting of 50 to 500 amino acid residues containing Z domain of Protein A or a fragment thereof, or a variant thereof, provided that the R binds to C-terminus or N-terminus of the R2.

Abstract: The present invention relates to immunoglobulin (Ig)-binding proteins with alkali-resistance properties. In one embodiment, the present invention provides for a variant of an Ig-binding protein, the variant comprising the Ig-binding protein having at least one asparagine residue substituted with a histidine, a serine, an aspartic acid or a threonine residue. The at least one substitution may confer to the variant Ig-binding protein an increased stability in alkaline solutions when compared to the wild-type Ig-binding. The present invention relates also to matrices for affinity separation of immunoglobulins comprising the Ig-binding proteins of the present invention, and to methods of using the Ig-binding proteins of the present invention to separate immunoglobulins from mixture compositions.

Abstract: Provided are a support for affinity chromatography which has excellent alkali resistance, and a method for isolating immunoglobulin. A support for affinity chromatography, containing an immobilized protein ligand represented by the following formula (1): R—R2??(1) wherein R represents a polypeptide consisting of 4 to 30 amino acid residues that contains an amino acid sequence represented by ATK or ASK; and R2 represents a polypeptide consisting of 50 to 500 amino acid residues containing an immunoglobulin-binding domain consisting of an amino acid sequence represented by SEQ ID NO: 1 or SEQ ID NO: 2, the partial sequence thereof, or an amino acid sequence having 70% or more identity to these sequences; with the proviso that a terminus at which R2 binds to R is C-terminus or N-terminus of the immunoglobulin-binding domain.

Abstract: The present invention relates to immunoglobulin (Ig)-binding proteins with alkali-resistance properties. In one embodiment, the present invention provides for a variant of an Ig-binding protein, the variant comprising the Ig-binding protein having at least one asparagine residue substituted with a histidine, a serine, an aspartic acid or a threonine residue. The at least one substitution may confer to the variant Ig-binding protein an increased stability in alkaline solutions when compared to the wild-type Ig-binding. The present invention relates also to matrices for affinity separation of immunoglobulins comprising the Ig-binding proteins of the present invention, and to methods of using the Ig-binding proteins of the present invention to separate immunoglobulins from mixture compositions.

Abstract: Provided are a support for affinity chromatography which has excellent alkali resistance, and a method for isolating immunoglobulin. A support for affinity chromatography, containing an immobilized protein ligand represented by the following formula (1): R—R2??(1) wherein R represents a polypeptide consisting of 4 to 30 amino acid residues that contains an amino acid sequence represented by ATK or ASK; and R2 represents a polypeptide consisting of 50 to 500 amino acid residues containing an immunoglobulin-binding domain consisting of an amino acid sequence represented by SEQ ID NO: 1 or SEQ ID NO: 2, the partial sequence thereof, or an amino acid sequence having 70% or more identity to these sequences; with the proviso that a terminus at which R2 binds to R is C-terminus or N-terminus of the immunoglobulin-binding domain.

Abstract: 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.

Abstract: Provided are a filler for affinity chromatography which has excellent alkali resistance, and a method for isolating immunoglobulin. The filler for affinity chromatography is a filler in which a protein represented by the following formula (1) is immobilized on a carrier. R—R2 ??(1) wherein R represents an amino acid sequence consisting of 4 to 300 amino acid residues containing a region consisting of 4 to 20 contiguous histidine residues; and R2 represents an amino acid sequence capable of binding to immunoglobulin, the amino acid sequence consisting of 50 to 500 amino acid residues containing Z domain of Protein A or a fragment thereof, or a variant thereof, provided that the R binds to C-terminus or N-terminus of the R2.

Abstract: Provided is a filler for affinity chromatography which is useful for protein purification and contains porous particles that have a high dynamic binding capacity for proteins and excellent pressure characteristics. The filler for affinity chromatography of the present invention is characterized in that it includes a porous particle consisting of a polymer of vinyl monomer including a cross-linkable vinyl monomer that contains hydroxyl group but does not contain epoxy group and an epoxy group-containing non-cross-linkable vinyl monomer, or a cross-linkable vinyl monomer that contains hydroxyl group and epoxy group, ligands bound to the porous particle, and ring-opened epoxy groups.