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Anal Biochem. 1991 Aug 15;197(1):137-42.

Internally standardized amino acid analysis for determining peptide/carrier protein coupling ratio.

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Center for Advanced Biotechnology and Medicine, Piscataway, New Jersey 08854.


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.

[Indexed for MEDLINE]

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