Development of High-Throughput ELISA Based Binding Assays to Detect p53/p63/p73 Family Protein-DNA Interaction in the 96-well Microplate Format for Drug Screening and Other Clinical and Diagnostic Uses
This technology includes ELISA based binding assays of p53, p63 or p73 provide possibilities to validate genome sequencing results, and allow the performance of more in-depth investigation to address scientific mechanisms, as well as to develop applications for high-throughput clinical and diagnosis usages. While quantitative p53 binding assays have been commercially developed, there is a lack of high-throughput method to detect binding activity of all three p53/p63/p73 family members, which are an important step for these transcription factors to perform their function. Targeting these transcription factors is an important strategy for drug development or clinical diagnostics. These binding assays will accelerate such process. Recent technology development of high-throughput sequencing has revealed genome-wide abnormality of p63/p73 binding in cancers. However, there is a lack of a high-throughput technology to validate the results generated from such technology, such as ChIP-seq (Chromatin Immunoprecipitation-parallel sequencing). This technology bridges the gap and provides the necessary validation.
- The binding assays for p53, p63 and p73 transcription factors could be used for drug screening of small molecule inhibitors.
- This invention can be developed as a commercially available kit for detecting the binding activities of NF-kB transcription factors. p53, p63 p73 binding assays can be commercialized and developed into similar kits to meet the needs for the research community.
- Currently, study of transcription binding activities is dependent on conventional methods such as EMSA (electrophoretic mobility shift assay), or ChIP (Chromatin Immunoprecipitation), which are very time consuming, and not suitable for high throughput application.
- The EMSA method needs to use radioisotopes for high sensitivity and reliability, which is not environmentally friendly.
- There is a lack of high-throughput method to validate lots of results from high throughput ChIP-seq or drug screening. However, these kits can be used for the high-throughput experiments, either in the academics or commercially.