| 摘要 |
An object of the present invention is to provide a carrier on which Protein A has been immobilized, the Protein A having a specific amino acid sequence that permits the desorption of antibodies under mild pH conditions (specifically, pH 4.0 to 5.5) in which many antibodies do not undergo acid modification; and a manufacturing method for the carrier. An immobilization carrier (excluding an immobilization carrier having a monolith structure) on which a protein is adsorbed by electrostatic interaction, the protein consisting of an amino acid sequence represented by the general formula: R1-R2-R3-R4-R5-R6, wherein the part represented by R1-R2-R3 is used for immobilization on the immobilization carrier, wherein: the sequence represents a sequence from the amino-terminal side towards the carboxyl-terminal side; the sequence of the R2 part is the sequence of a Protein A mutant as a protein to be immobilized or a sequence in which 1 to 3 units of the sequence thereof were linked together, the Protein A mutant having characteristics of strongly binding to an antibody under neutral conditions and dissociating with the antibody bound under neutral conditions under weakly acidic conditions of pH 4.0 to 5.5. |
| 主权项 |
1. An immobilization carrier, excluding an immobilization carrier having a monolith structure, onto which a protein is adsorbed by electrostatic interaction, the protein consisting of an amino acid sequence of the general formula: R1-R2-R3-R4-R5-R6, wherein R1-R2-R3 is used for immobilization on the immobilization carrier, wherein:
the amino acid sequence is a sequence from the amino-terminal side towards the carboxyl-terminal side; R1 may be absent, and, if present, is Met; R2 is the sequence of a mutant protein of the A-domain of Protein A consisting of a sequence selected from SEQ ID NOS: 4 to 7, or a sequence wherein 1 to 3 units of the sequence of the mutant protein of the A-domain of the Protein A consisting of a sequence selected from of SEQ ID NO: 4 to 7 are linked together; R3 may be absent, and, if present, is a spacer sequence consisting of 1 to 10 glycine residues; R4 is a sequence consisting of 2 residues of amino acids: cysteine-X, wherein X is alanine or an amino acid residue other than alanine and cysteine; R5 may be absent, and, if present, is a sequence of 1 to 10 amino acid residues consisting of aspartic acid, glutamic acid, or a combination thereof capable of rendering the isoelectric point of the protein consisting of the amino acid sequence of the general formula: R1-R2-R3-R4-R5-R6 on the acidic side; and R6 is an affinity tag sequence for the purification of proteins. |
