Description of Invention:
Gene expression profiling at the mRNA level has proven to be a powerful and useful tool, however this approach suffers from inherent limitations: 1) the mRNA abundance does not typically correlate well with protein abundance and 2) protein structure, activity, and function can be altered and regulated by post-translational modifications. Thus, there is growing recognition that these approaches should be complemented by profiles of the gene products or proteins themselves. The present invention provides methods for constructing and using a novel Monoclonal Antibody Microarray which allows high-throughput determination of protein expression profiles from serum, tissue, and cultured cells.
The Monoclonal Antibody Microarray consists of more than 1000 different antibodies immobilized on a glass slide, which recognize antigens from several groups of proteins, including cytokines, kinases, apoptotic proteins, growth factor receptors, tumor suppressors, and oncoproteins. Protein samples to be identified and quantified are labeled with fluorescence and hybridized to the antibodies immobilized on the arrays. By differentially labeling two protein samples (dual-color labeling) and co-hybridizing to the same microarray, a direct comparative analysis of protein expression can be performed using as little as 100 ìg of total protein. This method allows a large number of samples to be screened in parallel on identical arrays.
Applications:
High-throughput analysis of protein expression
Direct measurement of protein expression at the gene product or post-translational levels
Development Status:
The microarrays' performance was tested by proteomic profiling of two NCI-60 cancer cell lines (Renal UO-31 and Leukemia HL-60), demonstrating a high level of reproducibility.
The microarrays' performance was further evaluated by analysis of the protein expression profiles of 12 Borderline ovarian and 9 Adenocarcinoma ovarian tumors using normal ovarian surface epithelial cells as a reference cell line. It was possible to detect 77 proteins that showed statistically significant (p<0.05) differences distinguishing Borderline tumors and Adenocarcinoma tumors, demonstrating that the novel microarrays described are useful tools for proteomics.
Inventors:
Cassio S. Baptista (NCI) Lionel Best (NCI) David J. Munroe (NCI)
Patent Status:
DHHS Reference No. E-207-2006/0 --
U.S. Provisional Application No. 60/797,301 filed 02 May 2006
Licensing Status: Available for non-exclusive or exclusive licensing.
Collaborative Research Opportunity:
The NCI-Laboratory of Molecular Technology is seeking statements of capability or interest from parties interested in collaborative research to further develop, evaluate, or commercialize this novel monoclonal antibody microarray. Please contact Betty Tong, Ph.D. at 301-594-4263 or tongb@mail.nih.gov for more information.
Portfolios: Devices/Instrumentation Cancer
Cancer -Diagnostics-In Vitro-MAb Based Cancer -Diagnostics Devices/Instrumentation-Diagnostics Devices/Instrumentation-Research Materials
For Additional Information Please Contact: Cristina Thalhammer-Reyero PhD MBA
NIH Office of Technology Transfer
6011 Executive Blvd, Suite 325
Rockville, MD 20852-3804
Phone: (301) 435-4507
Email: thalhamc@mail.nih.gov
Fax: (301) 402-0220
