Journal of Proteomics & Bioinformatics
Open Access
ISSN: 0974-276X
+44 1223 790975
ISSN: 0974-276X
+44 1223 790975
Amita Nand, Deb K. Chatterjee and Zhu Jingsong
Posters: J Proteomics Bioinform
In recent years, in situ protein microarray unveils new methods to enable protein array to be created on demand. These methods exists by the use of cell free protein expression systems to produce proteins directly onto an immobilizing surface from prearrayed DNA or RNA and addressing the issues of laborious process and integral cost of protein expression and purification. Among these, various techniques have been developed such as NAPPA (nucleic acid programmable protein array) and TUS-TER microarray. In this research we are utilizing TUS-TER microarray technology to allow non-labeled detection with real time kinetics of protein-protein interactions through Surface Plasmon Resonance Imaging (SPRi). We constructed an expressionready plasmid DNA to generate protein arrays that serve dual function for synthesis of the protein of interest and function of capturing the expressed protein on the array with high binding affinity of TUS to TER (~3?7�10−13M). Plasmid DNA was immobilized on the amino modified gold surface and coupled with cell free expression system. All this process was performed in situ on SPRi within the flow cell. Furthermore, in situ expression and capturing of protein was confirmed by protein specific antibody. The current study develops a new method for in situ protein expression microarray with low cost and has potential to study the kinetics of protein-protein interaction. Taken together our findings, a protein microarray can be created as and when required without the problems of availability and stability associated with protein arrays for practical applications of Drug Discovery and Biomarker Discovery.
Amita Nand has completed his master degree from Amity University, India and recently he is doing Ph.D at National center for Nanoscience and Technology, Beijing. His research area focuses on protein-protein interaction and in situ protein microarray synthesis.