This invention relates to a hybrid ligand, a hybrid biomimetic chromedia and a preparing method and a use thereof, wherein the hybrid biomimetic chromedia takes hydrophilic porous microsphere as a substrate in chromatography, activated with allyl bromide and undergoing bromo-alcoholization with N-bromosuccinimide, then coupled with the hybrid ligands. The sequence of the hybrid ligand is phenylalanine-tyrosine-glutamine-5-aminobenzimidazole. The hybrid biomimetic chromedia has both of the two functional groups of phenylalanine-tyrosine-glutamine tripeptide and aminobenzimidazole, while maintaining the high antibody selectivity of polypeptide ligand, hydrophobic electric charge inductive ligand is introduced to achieve more moderate elution requirement, realizing effective antibody separation.

The present invention relates to an immunoglobulin-binding protein, wherein at least one asparagine residue has been mutated to an amino acid other than glutamine or aspartic acid, which mutation confers an increased chemical stability at pH-values of up to about 13-14 compared to the parental molecule. The protein can for example be derived from a protein capable of binding to other regions of the immunoglobulin molecule than the complementarity determining regions (CDR), such as protein A, and preferably the B-domain of Staphylococcal protein A. The invention also relates to a matrix for affinity separation, which comprises an immunoglobulin-binding protein as ligand coupled to a solid support, in which protein ligand at least one asparagine residue has been mutated to an amino acid other than glutamine.

The present invention relates to an immunoglobulin-binding protein, wherein at least one asparagine residue has been mutated to an amino acid other than glutamine or aspartic acid, which mutation confers an increased chemical stability at pH-values of up to about 13-14 compared to the parental molecule. The protein can for example be derived from a protein capable of binding to other regions of the immunoglobulin molecule than the complementarity determining regions (CDR), such as protein A, and preferably the B-domain of Staphylococcal protein A. The invention also relates to a matrix for affinity separation, which comprises an immunoglobulin-binding protein as ligand coupled to a solid support, in which protein ligand at least one asparagine residue has been mutated to an amino acid other than glutamine.

An Fc-binding polypeptide of improved alkali stability, comprising a mutant of an Fc-binding domain of Staphylococcus Protein A (SpA), as defined by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO:3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO 26 or SEQ ID NO 27, wherein at least the alanine residue at the position corresponding to position 42 in SEQ ID NO:4-7 has been mutated to arginine and/or wherein at least the aspartic acid residue at the position corresponding to position 37 in SEQ ID NO:4-7 has been mutated to glutamic acid.

An Fc-binding polypeptide of improved alkali stability, comprising a mutant of an Fc-binding domain of Staphylococcus Protein A (SpA), as defined by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO:3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO 26 or SEQ ID NO 27, wherein at least the alanine residue at the position corresponding to position 42 in SEQ ID NO:4-7 has been mutated to arginine and/or wherein at least the aspartic acid residue at the position corresponding to position 37 in SEQ ID NO:4-7 has been mutated to glutamic acid.

Problems to be Solved The present invention provides an affinity support capable of trapping a substance by cooperative binding that is less likely to cause dissociation even when the substance is a molecule other than an antibody, and a trapping method using the same. Means to Solve the Problems A method of trapping a substance comprising the step of contacting an objective to be trapped with an affinity support comprising a support, a spacer bound to the support and an affinity substance bound to the spacer, so as to bind the objective to be trapped to the affinity substance, wherein each one of the objective to be trapped has a plural of affinity sites and the affinity substance binds to at least two of the affinity sites simultaneously.

Problems to be Solved The present invention provides an affinity support capable of trapping a substance by cooperative binding that is less likely to cause dissociation even when the substance is a molecule other than an antibody, and a trapping method using the same. Means to Solve the Problems A method of trapping a substance comprising the step of contacting an objective to be trapped with an affinity support comprising a support, a spacer bound to the support and an affinity substance bound to the spacer, so as to bind the objective to be trapped to the affinity substance, wherein each one of the objective to be trapped has a plural of affinity sites and the affinity substance binds to at least two of the affinity sites simultaneously.

One embodiment of the present invention is a protein having affinity for an immunoglobulin, which is a protein having two or more domains derived from any of the amino acid sequences of E, D, and A domains of protein A, and in the amino acid sequence of at least one of the domains, one or more lysines are included, and the C-terminal lysine is deleted or substituted, or a protein having affinity for an immunoglobulin, which is a protein having two or more domains derived from any of B, C, and Z domains of protein A, and in the amino acid sequence of at least one of the domains, one or more lysines are included, and lysine at position 4 and the C-terminal lysine are deleted or substituted.

One embodiment of the present invention is a protein having affinity for an immunoglobulin, which is a protein having two or more domains derived from any of the amino acid sequences of E, D, and A domains of protein A, and in the amino acid sequence of at least one of the domains, one or more lysines are included, and the C-terminal lysine is deleted or substituted, or a protein having affinity for an immunoglobulin, which is a protein having two or more domains derived from any of B, C, and Z domains of protein A, and in the amino acid sequence of at least one of the domains, one or more lysines are included, and lysine at position 4 and the C-terminal lysine are deleted or substituted.

Adsorbent particles including: porous carrier particles containing an organic polymer; polyethylenimine attached to a surface of the porous carrier particles; and a diglycolic acid residue bonded to an amino group of the polyethylenimine. A most frequent pore size of the porous carrier particles exceeds 10 nm.