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Effects of histidine residues on adsorption equilibrium of peptide antibodies
Histidine and arginine residues in a peptide, which represent residues 112-121 of myoglobin, were replaced by alanine, and antibodies for these peptides were raised in rabbits. Adsorption equilibria between these peptides and antibodies were measured to identify the essential amino acid residues for specific binding and their effects on adsorption behavior. Protonation of one histidine residue affected strongly the dependence of adsorption equilibrium on pH, and the adsorption capacities of the antibodies against the peptides containing this essential histidine residue rapidly decreased at around pH 5-6, which corresponds to the pKa value of histidine. Immunization with peptide sequences containing such histidine residues is one method to obtain suitable antibodies for immunoaffinity chromatography, which have a weak affinity for the elution step under mild conditions.
Histidine and arginine residues in a peptide, which represent residues 112-121 of myoglobin, were replaced by alanine, and antibodies for these peptides were raised in rabbits. Adsorption equilibria between these peptides and antibodies were measured to identify the essential amino acid residues for specific binding and their effects on adsorption behavior. Protonation of one histidine residue affected strongly the dependence of adsorption equilibrium on pH, and the adsorption capacities of the antibodies against the peptides containing this essential histidine residue rapidly decreased at around pH 5-6, which corresponds to the pKa value of histidine. Immunization with peptide sequences containing such histidine residues is one method to obtain suitable antibodies for immunoaffinity chromatography, which have a weak affinity for the elution step under mild conditions.