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Design and Functional Analysis of ssDNA Directed Assembly in Protein Array

Design and Functional Analysis of ssDNA Directed Assembly in Protein Array content piece image

Abstract
In the post genomic era, the characterization of complex cellular functions requires the large scale analysis of the proteome. Antibody microarray technology has therefore become an invaluable tool in understanding the level of protein expression as well as protein-protein interaction in a high throughput manner. However, current fabrication approach of antibody microarray often results in the loss of antibody functionalities after immobilization onto the substrate.

The present poster discusses the fabrication and evaluation of a new platform called “Spatially addressable protein array” (SAPA). By exploring the specificity of DNA hybridization, ssDNA-antibody conjugates would capture the antigen from complex biological samples in milieu and spatially addressed to specific location on the oligo array for detection.

Such approach allows the protein-analyte interaction to take place in a solution phase and reduce the loss of antibody functionality due to unfavorable surface protein interactions. Further optimization has been done by investigating surface chemistry, non-specific protein adsorption and facile preparation of the ssDNA-conjugated antibody. Experimental studies have shown that the platform is able to detect samples at pM scale and it could be fine-tuned to achieve an optimal system for solving biological problems.