Accession Number : ADA333262

Title :   Development of Anti-Idiotype Monoclonal Antibodies for the Treatment of Breast Cancer

Descriptive Note : Final rept 1 Jul 94-30 Jun 97

Corporate Author : KENTUCKY UNIV LEXINGTON

Personal Author(s) : Chatterjee, Malaya

PDF Url : ADA333262

Report Date : JUL 1997

Pagination or Media Count : 18

Abstract : For the development of HER2/neu mimicking anti-idiotype antibodies (Ab2s), groups of female BALD/c mice were immunized with three sets of monoclonal antibodies (Ab1), designated 520C9, 741F8 and 454C11, directed at distinct antigenic determinants of HER2/neu. Hybridomas were generated from each treatment group and one, stable Ab2 producing hybrid each of 520C9 and 741F8 were obtained, and cloned twice by limiting dilution. The isotypes of 520C9 and 741F8 Ab2's were IgG1k by ELISA. 520C9 and 741F8 Ab2 cells were used to produce mouse ascites and the Ab2 purified by affinity chromatography and confirmed by SDS-PAGE. A competitive binding assay using Ab2, Ab1 and the human breast cancer cell line SK-BR-3 (which express HER2/neu on their cell surface), showed that 50mu1 of each of 520C9 and 741F8 Ab2 containing supernatants inhibited the binding of Ab1 to SK-BR-3 cells by tilde 80%.The specificity binding study showed that the binding of 741F8 Ab2 was to those antibodies directed against HER/2neu. A competitive binding assay using 741F8 Ab2, Ab1 and SK-BR-3 cells showed that 10ng of purified 741F8 Ab2 inhibited the binding of Ab1 to SK-BR-3 cells by tilde 50%. Polyclonal anti-anti-idiotype antibodies (Ab3), were generated in rabbits using either purified 520C9 Ab2 or 741F8 Ab2. The Ab3 reacted with HER2/neu positive tumor cells suggesting the presence of anti-tumor (Ab1') antibodies.

Descriptors :   *MONOCLONAL ANTIBODIES, *BREAST CANCER, HUMANS, NEOPLASMS, CLONES, IMMUNOLOGY, CELLS(BIOLOGY), ASSAYING, ANTIGENS, FEMALES, CHROMATOGRAPHY, ASCITES, HYBRIDOMAS.

Subject Categories : Biochemistry
      Medicine and Medical Research
      Pharmacology

Distribution Statement : APPROVED FOR PUBLIC RELEASE