ANTIGENIC STRUCTURE OF PROLINE-RICH REGION-536-566 OF PERTACTIN FROM BORDETELLA-PERTUSSIS .1. PREPARATION AND ANALYSIS OF ANTISERA AGAINST SYNTHETIC PEPTIDES

Rabbit antisera were raised against four synthetic peptides from the P ro-rich epitope of pertactin from B. pertussis. Prior to immunization peptides KAPPAPKPAPQPGPQPP (17P), QPPQPQPEAPAPQP (14P), GPQPPQPPQP (10 P), and PGPQPP (6P) were conjugated to keyhole limpet hemocyanin (KLH) . The highest titers of antibodies against the synthetic peptides as m easured by ELISA were 1:4,000,000 (17P), 1:64,000 (14P), and 1:256,000 (10P). The antibodies raised against KLH-6P did not react with 6P-coa ted ELISA plates, but recognized the hexapeptide conjugated to ovalbum in. Indirect and competitive ELISAs demonstrated that none of the anti sera against the conjugates (KLH-17P, KLH-14P, KLH-10P, and KLH-6P) cr oss-reacted with unrelated antigens. Antisera against peptides 10P, 14 P, and 17P and anti-peptide antibodies purified using affinity chromat ography differed in their ability to recognize the intact pertactin.