UNIT 11.16 Synthetic Peptides for Production of Antibodies that Recognize Intact Proteins
Published Online: 1 AUG 2002
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Grant, G. A. 2002. Synthetic Peptides for Production of Antibodies that Recognize Intact Proteins. Current Protocols in Molecular Biology. 59:IV:11.16:11.16.1–11.16.19.
- Published Online: 1 AUG 2002
- Published Print: JUL 2002
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.