ovalbumin and 2-aminoisobutyric-acid

A method based on amino acid analysis has been developed for monitoring the covalent conjugation of synthetic peptide haptens to carrier proteins. The marker amino acid, alpha-aminobutyric acid, is included in the sequence during peptide synthesis. Following reaction, the carrier protein-conjugate is freed of excess peptide by two successive rounds of gel filtration chromatography. Amino acid analysis of a hydrolysate of the conjugate allows the calculation of the coupling ratio of the peptide to the carrier protein. Two typical procedures for conjugation, carbodiimide cross-linking and cysteine-thiol reaction with maleimidyl-proteins, have been evaluated.

Topics: Amino Acid Sequence; Amino Acids; Aminoisobutyric Acids; Carrier Proteins; Chromatography, Gel; Cysteine; Ethyldimethylaminopropyl Carbodiimide; Hemocyanins; Molecular Sequence Data; Ovalbumin; Peptides; Reference Standards