Antibody Microchips to Study Metastasis
Abstract
The hypothesis of my project is the behavior of an invasive tumor cell is largely determined by a collection of proteins on the surface of metastasis cell. I proposed to employ phage display single-chain variable fragment antibody library, mass spectrometry and antibody chip methodologies to study the molecular mechanisms of metastasis in breast carcinoma MDA-MB-435 cell line. During the first year of this grant, I have constructed a phage display scFv antibody library to the plasma membrane proteins of this cell line. I panned the library on intact cells to enrich population of phage-scFvs that are composed of antibodies to cell surface proteins of MDA-MB-435 cell, 5.6-fold, 14-fold and 20-fold enrichment was obtained as compared with background binding when three rounds subsequent pannings were performed. The specificity of 705 individual phage clones was tested by an ELISA assay. Forty-six of seven hundred and five phage clones showed 2.5-fold greater specificity to MDA-MB-435 cells as compared with normal mammary epithelial cells. Ten of these clones bind to MDA-MB-435 tumor cells in flow cytometry assays. These clones are being sub-cloned in preparation for expression as soluble scFv antibodies. The recombinant scFvs will be used to identify the target antigens.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 2002
- Accession Number
- ADA409806
Entities
People
- Yan Zhang
Organizations
- Sanford Burnham Prebys Medical Discovery Institute