Development of Antibody Microarrays for the National Cancer Institute EDRN

National Cancer Institute, Frederick MD

Investigators

Abstract

Stage I: A feasibility study will require the generation of sandwich Enzyme-Linked Immunosorbent Assay (ELISA) like microarray composed of immobilized scFv antibodies, 5 different affinities for each of the two antigens, EGF and PSA, as well as comparison to commercial monoclonal and polyclonal antibodies specific for these antigens. The assay will require additional detection antibodies directed at a different epitope(s). Those will be composed of high affinity scFv antibodies against PSA and EGF respectively and commercial detection antibodies (monoclonal or polyclonal). This platform should test the performance of scFv antibodies versus other monoclonal and polyclonal antibodies. In addition, the platform should perform equally well or better than a commercial sandwich ELISA in quantitative detection of these antigens for human serum or plasma. The utility of the scFv antibodies as affinity reagents for immunohistochemistry (IHC) should be tested in collaboration with an EDRN investigator. Stage II: Similar to Stage I except that it will be limited to approximately 50 antigens (markers) and controls Stage III: Validation of the technology using clinical specimens and controls - In this stage PNNL will train selected EDRN investigators on how to use the microarray platform, print a large number of microarrays for analytical and clinical validation studies