EGFR-Specific Single-Chain Antibody-Taxol Conjugate for Radiosensitizing Breast Carcinomas.

摘要

We propose to conjugate paclitaxel to a single-chain antibody that specifically binds to the extracellular domain of the EGFR. The scFv-paclitaxel conjugate will be able to localize to the tumor rapidly, penetrate deeper, be eliminated quickly, and induce fewer side effects for the patient, thereby increasing the efficacy of the scFv-paclitaxel conjugate. An additional benefit with the scFv-paclitaxel conjugate is the fact that taxol is a radiosensitizing drug, therefore radiation treatment can be used following the administration of the scFv-paclitaxel conjugate. The EGFR overexpressing breast cancer cell line, MDA-MB-468 will be used in this proposal to show 'proof of concept'. Cell proliferation assays will be used to examine the cytotoxicity of the scFv- paclitaxel conjugate as compared to paclitaxel, scFv or the combination of the two agents. Cell death will be examined using two different apoptosis assays, annexin V-FITC and TUNEL. Since taxol is a radio sensitizing drug, radiation will be included in the cell proliferation and apoptosis assays. Clonogenic assays will be done to determine the radio sensitizing ability of the scFv- paclitaxel conjugate as compare to each agent alone.