A Parallel High-Throughput Pharmacogenetic Profiling Assay for Clinical Use

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DESCRIPTION (provided by applicant): A Parallel High Throughput Pharmacogenetic Profiling Assay for Clinical Use Pharmacogenomic studies have already identified a number of genes whose SNP genotypes or haplotypes correlate with different individual drug responses, other metabolic processes or disease susceptibility. Thus the ability to determine genotypes quickly and accurately for medically relevant regions is critical to understanding the effects of an individual's genetic profile on these processes, and to the development of predictive, preventative and personalized medicine. Maxwell Sensors Inc. proposes to develop Barcode Etched Assayable Micro Bead (BEAM bead) technology, which combines digital barcoded beads and molecular chemistry for a pharmacogenetic SNP genotyping assay, which can be used for the high throughput molecular profiling of individuals. The digital BEAM beads are paramagnetic and are fabricated by photolithography to have thousands of identification codes and can be functionalized with nucleic acids, proteins or other small molecules to carry out large multiplexed assays in homogeneous or heterogeneous media. Genotyping using BEAM beads offers the advantages of flexibility, high throughput, easy fabrication, low cost mass production, and all in one reaction in small volumes. During Phase I of the project, we have successfully (1) fabricated 10 digit BEAM beads, allowing 1,024 (=210) unique codes, making it the most highly multiplexed barcode magnetic bead in the world; (2) designed and constructed a microfluidic transducer and fluorescence detector to decode and detect fluorescence one by one; (3) developed multiplex PCR for P450 (CYP2A7), KCNA5 and ITPase, and bead hybridization; and (4) evaluated the SNP genotyping assay using multiplex PCR amplified sequences. Phase II work will focus on optimization of the BEAM bead platform for clinical applications, design and integration of BEAM system, design of standard and custom SNP panels for genotyping SNPs relevant to drug responses, and development of multiplex PCR for target gene amplification and SBE hybridization.