Production of rabbit antibody specific for amino-terminal residues of cholecystokinin octapeptide (CCK-8) by selective suppression of cross-reactive antibody response.
Antibody specific for the amino-terminal region of cholecystokinin octapeptide (CCK-8) was generated in a highly reproducible way in New Zealand white rabbits by a novel immunization procedure which involves immunization with CCK-8 peptide conjugate coupled with keyhole limpet hemocyanin (KLH) and inhibiting cross-reacting antibody formation by treatment of the animals with a potent tolerogenic conjugate of beta-alanyl-tetragastrin and a copolymer of D-glutamic acid and D-lysine (D-GL). The antisera thus produced specifically react with an amino-terminal region of CCK-8 but not with the non-sulfate form of CCK-8, nor with the carboxy-terminal region which shares a cross-reactive determinant among gastrin and cholecystokinin-related peptides (caerulein, CCK-4, CCK-8, CCK-33 and CCK-39). The antisera produced by this method allowed us to measure specifically CCK in extracts from tissue such as duodenum containing gastrin and CCK at comparable levels.
D000596:Amino Acids / Q000276:immunology*; D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D001923:Brain Chemistry; D002766:Cholecystokinin / Q000276:immunology*; D003429:Cross Reactions; D004386:Duodenum / Q000032:analysis; D005755:Gastrins / Q000032:analysis / Q000494:pharmacology; D006801:Humans; D007106:Immune Sera / Q000032:analysis / Q000494:pharmacology; D007158:Immunologic Techniques; D010446:Peptide Fragments / Q000276:immunology*; D011817:Rabbits; D051381:Rats; D012844:Sincalide; D013552:Swine
D000596:Amino Acids; D005755:Gastrins; D007106:Immune Sera; D010446:Peptide Fragments; C008530:minigastrin; D002766:Cholecystokinin; D012844:Sincalide