Generation of an antibody with a designed specificity difference for protein C and activated protein C.
Rabbit polyclonal antibodies to a synthetic peptide, NH2-Asp-Thr-Asn-Gln-Val-Asp-Gln-Lys-Asp-Gln-Leu-Asp-Phe-Arg-CONH2 (A Pep), have been produced. This sequence is identical to that contained in the tetradecapeptide released from bovine protein C (PC) as a result of its conversion to its activated form (APC), except that Phe13 replaced the normal Pro13, in order to discourage cross-reactivity of antibodies to the carboxylterminal portion of APep with PC. The antibody pool obtained reacted with PC and showed virtually no cross-reactivity toward either APC or several typical plasma proteins. This general approach should serve well as a means of production of antibodies with a designed specificity capable of distinguishing between forms of the same protein that arise by release of peptide material.
Zhang
L
L
Department of Chemistry, University of Notre Dame, Indiana 46556.
Castellino
F J
FJ
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology*; D011486:Protein C / Q000276:immunology*; D011817:Rabbits
D000596:Amino Acids; D010446:Peptide Fragments; D011486:Protein C