Accession Number:

ADA394199

Title:

Targeted Gene Therapy for Breast Cancer

Descriptive Note:

Final rept. 1 Aug 1997-31 Jul 2000

Corporate Author:

UNIVERSITY OF SOUTHERN CALIFORNIA LOS ANGELES

Personal Author(s):

Report Date:

2000-08-01

Pagination or Media Count:

36.0

Abstract:

The HER-2neu oncogene is amplified and overexpressed in 25-30 of patients with breast cancer. The gene encodes a protein receptor found on the surface of breast cancer cells in this subset of patients. Four novel monoclonal antibodies, 5A7, 8H11, 11F11, and 10H8, were isolated and characterized in this work. The ultimate aim of this work was to develop target specific gene therapy for breast cancer. To further that aim, we used a rapid radioimmunoassay to detect antibodies which could bind to live human breast cancer cells and trigger internalization into those cells. Monoclonal antibodies 8H11 and 10H8 were found to have this specific property. The next major step was the cloning of the variable region gene sequences of these monoclonal antibodies from the hybridoma cell lines isolated for antibody production. The ability to design degenerate primers for specific amplification of the DNA encoding these variable regions allowed for the cloning, sequencing, and then splicing of the variable heavy VH with the variable light VL genes. The subsequent single chain variable fragment scFv was cloned and determined to approximate the antigen binding site of the parent immunoglobulin. An attempt to characterize the protein product of the scFv gene of 10118 met with problems during protein expression. Therefore, the final aim of establishing target specificity to retroviral vectors ended with an unsuccessful endpoint which exceeded the ability of the trainee to complete in this funding period.

Subject Categories:

  • Biochemistry
  • Genetic Engineering and Molecular Biology
  • Medicine and Medical Research

Distribution Statement:

APPROVED FOR PUBLIC RELEASE