A novel design of peptide immunogens: synthetic peptide with a reversible handle for the attachment to protein carriers.
A novel design of peptide antigen suitable for the preparation of peptide-based antibody is described. The peptide is synthesized in a special benzhydrylamine (p-acyloxybenzhydrylamine) resin that contains two detachable sites, both of which are stable to the mild SN2 deprotection of the side chains that allows the deprotected peptide to remain attached to the resin support. Release of the peptide is achieved by methylaminolysis in a volatile solvent that produces a deprotected peptide with a detachable p-hydroxybenzhydrylamine handle for the linkage to the protein carrier via an azo bond. The peptidyl handle is detachable under a mild acidolytic condition to generate the desirable peptide. A 17-residue peptide from the circumsporozoite proteins of the human malaria parasite, Plasmodium flaciparum, was tested in this approach to prepare antibodies, and its results were compared with the conventional approach using glutaraldehyde as the crosslinking reagent. Specific antibodies obtained from this approach were found to be reactive with the peptide and the cognate protein. Our results show that this design is useful and convenient to generate site-specific peptide-based antibodies for laboratory use.
Vergara
U
U
Rockefeller University, New York, NY 10021.
Tam
J P
JP
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology; D000917:Antibody Formation; D000941:Antigens; D006801:Humans; D007202:Indicators and Reagents; D016778:Malaria, Falciparum / Q000276:immunology; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis* / Q000276:immunology; D010963:Plasmodium falciparum / Q000276:immunology; D011817:Rabbits / Q000276:immunology; D014614:Vaccines, Synthetic
D000913:Antibodies, Protozoan; D000941:Antigens; D007202:Indicators and Reagents; D010455:Peptides; D014614:Vaccines, Synthetic