Synthetic peptides for production of antibodies that recognize intact proteins

Grant, G.A.

Current Protocols in Molecular Biology Chapter 11: Unit 11.16

2002


ISSN/ISBN: 1934-3647
PMID: 18265297
DOI: 10.1002/0471142727.mb1116s59
Accession: 068853718

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Abstract
Antibodies that recognize intact proteins can be produced through the use of synthetic peptides based on short stretches of the protein sequence, without first having to isolate the protein. The procedure for selecting the antigenic sequence is relatively straightforward. This unit identifies ways to determine the best sequence and to chemically couple the synthetic peptide to a carrier protein to boost the immune response. Another method entails direct synthesis of a MAP covalent multimer of the simple peptide sequence. Peptides can also be coupled to carrier proteins through either their amino or carboxyl groups. Alternate protocols in this unit cover these scenarios. In addition, methods for assaying free sulfhydryl content and for reducing disulfide bonds in synthetic peptides are provided. Finally, once the coupling procedure has been performed, it is possible to determine the approximate degree of coupling by amino acid analysis, and a procedure for this is also provided.