PROCESS FOR IMMOBILIZING ORIENTATION-CONTROLLED PROTEIN AND PROCESS FOR ARRAYING AND IMMOBILIZING PROTEIN USING THE SAME

摘要

This invention provides a process for effectively producing an immobilized protein that is immobilized only at the carboxy terminus. This is a process for immobilizing on a carrier for immobilization a protein represented by general formula (1): <?in-line-formulae description="In-line Formulae" end="lead"?>NH2-R1-COOH (1)<?in-line-formulae description="In-line Formulae" end="tail"?> wherein a protein having a sulfhydryl group represented by general formula (2) is produced: <?in-line-formulae description="In-line Formulae" end="lead"?>NH2-R1-CONH-R2-CO-NH-CH(CH2-SH)-CO-NH-R3-COOH (2),<?in-line-formulae description="In-line Formulae" end="tail"?> the resulting protein is immobilized adsorptively on the carrier for immobilization represented by general formula (3) via ion interactions under neutral conditions: <?in-line-formulae description="In-line Formulae" end="lead"?>NH2-Y (3), and<?in-line-formulae description="In-line Formulae" end="tail"?> a sulfhydryl group of the cysteine residue in the protein represented by general formula (2) that is adsorbed on the carrier for immobilization represented by general formula (3) is cyanated using a cyanating reagent to convert it into a cyanocysteine residue, thereby producing an immobilized protein represented by general formula (4): <?in-line-formulae description="In-line Formulae" end="lead"?>NH2-R1-CO-NH-R2-CO-NH-Y (4)<?in-line-formulae description="In-line Formulae" end="tail"?> wherein R1 and R2 represent arbitrary amino acid sequences, R3 represents an amino acid sequence that has a strong negative charge around the neutral region and can acidify the isoelectric point of NH2-R1-CONH-R2-CO-NH-CH(CH2-SH)-CO-NH-R3-COOH, and Y represents an arbitrary carrier for immobilization.