International Symposium on LifeChips 2006

Functional Proteomics for Biomarker and Target Discovery
One of the most compelling steps in the post-genomic era will be learning the functional roles for all proteins. The Harvard Institute of Proteomics (HIP) has initiated a project to create a sequence-verified collection of full-length cDNAs representing all human coding regions in a recombinational vector system that allows the immediate in-frame transfer of all coding regions into virtually any protein expression vector. These transfers allow the addition of peptide tags to either or both end of the proteins. This repository, called the FLEXGene Repository (for Full-Length Expression-ready), will enable the high-throughput (HT) screening of protein function for the entire set (or any customized subset) of genes using any method of in vitro or in vivo expression. The most exciting part of this project has been the ease with which the clones from the repository can be rapidly incorporated in HT biological experimentation. Using automated gene transfer methods and protein purification it is now possible to purify thousands of proteins. Using HT retroviral methods, proteins capable of driving cell migration, altering the morphogenesis of normal epithelial structures, and affecting substrate dependent growth have been identified. A novel form of protein microarray, called nucleic acid programmable protein array (NAPPA), has been developed. This method substitutes the printing of proteins on the array with printing cDNAs encoding the proteins. Thus, the array is a DNA array that can be converted into a protein array by adding cell free protein synthesis machinery. This obviates the need to purify proteins, produces human proteins in a mammalian milieu, and avoids concerns about protein stability on the array because the proteins are made just-in-time for assay. NAPPA arrays can be used to study protein-protein interactions, protein-drug interactions and as tools to search for disease biomarkers.

Biography
This presentation will be given by Dr. Niroshan Ramachandran on behalf of Dr. LaBaer. Niroshan Ramachandran PhD is a research associate in the lab of Joshua LaBaer MD PhD at the Harvard Institute of Proteomics, Harvard Medical School. He has been actively involved in the development of protein microarray technology. This technology called Nucleic Acid Programmable Protein Array (NAPPA) (Science. 2004 305(5680),86-90), synthesizes an array of proteins on a solid matrix and overcomes many of the limitations of conventional protein arrays on the market today. He is currently developing this platform technology to be used for biomarker discovery in cancer. His goal is to identify informative antigens that will lead to early diagnosis of diseases and the development of vaccines for immunotherapy. Dr. Ramachandran received his Hon. BSc in 1997 from the University of Toronto, Canada. He performed his doctorate work in the area of nitric oxide metabolism in the lab of Dr. Bulent Mutus, University of Windsor, Canada. He joined the Harvard Institute of Proteomics as a post doctoral fellow in 2001.