Production of Anti-idiotypic Monoclonal Antibody Mimics for Coronatine

Abstract

Coronatine (COR) is composed of two structural components, coronafacic acid (CFA) and the amino acid coronamic acid (CMA), which are joined by an amide bond. Monoclonal anti-idiotypic antibodies were prepared and used in competitive ELISAs. Monoclonal antibodies (MAbs) were secreted by hybridoma cell lines prepared from mice immunized with two different idiotypic MAbs, 11B8 and 11G8 that predominantly recognized the CMA or CFA residues, respectively. Hybridoma cell lines secreting high affinity MAbs 4D5 (anti 11B8) and 4H10 (anti 11G8) were isolated. MAb 4D5 predominantly mimicked the coronamyl amide features of coronatine whereas 4H10 mimicked the coronafacoyl amide of COR. Immunoassays were developed using each of these antibodies. COR could be quantified in the range 1-1000 ngml m 1 (limit of detection <150 pgml m 1 ) in direct and modified indirect assays for 4D5/11B8 and in direct assays for 4H10/11G8. For modified indirect assays using 4H10/ 11G8 COR was quantified in the range 3-1000 ngml m 1 (limit of detection <1 ngml m 1 ).