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.
Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex® technology (see box on right).READ MORE
Genome-wide association studies (GWAS) have identified more than 100 genetic loci associated with type 2 diabetes. The majority of these are located in the intergenic or intragenic regions suggesting that the implicated variants may alter chromatin conformation. This, in turn, is likely to influence the expression of nearby or more remotely located genes to alter beta cell function. At present, however, detailed molecular and functional analyses are still lacking for most of these variants. We recently analysed one of these loci and mapped five causal variants in an islet-specific enhancer cluster within the STARD10 gene locus. Here, we aimed to understand how these causal variants influence b-cell function by alteration of the chromatin structure of enhancer clusterREAD MORE
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