Design and Functional Analysis of ssDNA Directed Assembly in Protein Array
Poster Feb 01, 2007
Ng Jin Kiata, Parayil Kumaran Ajikumara, Tang Yew Chunga, Lee Jim Yanga, Gregory Stephanopoulosa and Too Heng-Phona
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.
When there is a need to quickly analyze samples using a number of different PCR assays, it is likely that optimal conditions for each assay will not be the same. First, different assays often will require different annealing temperatures for their primers. In addition, amplicons may be designed to be of different lengths and therefore require varying durations of the extension step.READ MORE