Selectivity of angiotensin II antisera.
A significant problem in the immunoassay of angiotensin II is the cross-reactivity of most available antisera with the peptide's metabolic products, (des-Asp1)-angiotensin II and (des-Asp1.Arg2)-angiotensin II. In order to attempt to generate antisera of greater selectivity, a variety of conjugates between angiotensin II or derivative peptides and carrier proteins were examined as immunogens with the aim of generating antisera that would selectively identify the amino terminal region of the peptide. Selectivity for the amino terminus was achieved by either (1) immunization with N-acetylated angiotensin II-amide which had been coupled to rabbit serum albumin by its carboxy terminus, or (2) immunization with angiotensin-(1-7)-heptapeptide which was randomly coupled to thyroglobulin. The antisera produced with the N-acetylated immunogen cross-reacted with the unacetylated ligand (Asn1-Val5)-angiotensin, but did not recognize the human hormone (Asp1,Ile5)-angiotensin. Carboxy-terminal coupling of angiotensin without N-acetylation did not induce selectivity for the amino terminus, nor did a conjugate which was linked to the carrier protein via a diazo bond to His6 of the peptide. These findings may be explained by the fact that N-acetylated angiotensin II resists degradation by amino peptidases and thus retains its structure in the immunogen and by the fact that the (1-7)-heptapeptide has lost the immunodominant carboxy-terminal epitope, thus emphasizing the desired amino terminal determinant.
D000804:Angiotensin II / Q000032:analysis / Q000276:immunology*; D000818:Animals; D000915:Antibody Affinity; D002352:Carrier Proteins / Q000276:immunology; D002417:Cattle; D003429:Cross Reactions; D003979:Diazonium Compounds / Q000276:immunology; D007106:Immune Sera / Q000494:pharmacology; D007118:Immunoassay / Q000379:methods; D011817:Rabbits; D013954:Thyroglobulin / Q000276:immunology
D002352:Carrier Proteins; D003979:Diazonium Compounds; D007106:Immune Sera; D000804:Angiotensin II; D013954:Thyroglobulin