New Multiplexed Quantitative Detection of Pluripotency and Germ Layer Proteins
DESCRIPTION (provided by applicant): Primorigen Biosciences will use Phase II SBIR funds to accelerate and expand the development of new regenerative medicine cell therapies by developing a novel high throughput, low cost protein quantification system for characterizing pluripotent cell lines and early germ layer differentiation. Primorigen successfully confirmed the technical feasibility of this proposal by achieving both major goals of the Phase I project, first demonstrating (and now routinely using) its proprietary Spots on Dots frameless microarray platform for high-throughput antibody screening and characterization, and then demonstrating feasibility of developing a multiplexed protein quantitation assay for cell pluripotency by identifying matched pairs of antibodies (working sandwich assays) for four key pluripotency targets: Sox2, Oct4, Nanog and Rex1. In Phase II, Primorigen will expand the panel of antibodies for pluripotency targets and develop a second array consisting of antibodies for 9 markers of early germ layer differentiation. These combined tools will provide a more complete solution for tracking downstream development of pluripotent cells, and will be validated using a variety of appropriately differentiated stem cells, and by obtaining independent validation of the performance and specificity of the assays. In addition, Primorigen will collaborate with a software developer to produce a software/flatbed scanner package that enables standardized analysis and reporting of data from assays run on the Spots on Dots platform in a rapid, 'one touch' format. PUBLIC HEALTH RELEVANCE: Proteins facilitate many of the cell's most basic functions of reproduction, metabolism, growth, and programmed death. In the past few years, thousands of new protein sequences and post- translational variants have been identified by the human genome and proteome projects. This protein jackpot has hastened the drive to understand protein-based regulation but it also challenges researchers to find better biophysical methods to quantitatively detect protein markers. As a result, there is demand for inexpensive, miniaturized, multiplexed high throughput assays that can generate thousands of protein detection data points per experiment. Primorigen's SBIR proposal will address this need for the cell therapy research community by developing well characterized antibody and protein assay content for detecting and quantifying human cell markers of pluripotency and early germ layer differentiation, and then formatting these in a high throughput, low-cost multiplexed immunoassay format in a frameless microarray that is supported by one touch user friendly software for data analysis and reporting.
Small Business Information at Submission:
PRIMORIGEN BIOSCIENCES, INC.
510 CHARMANY DRIVE MADISON, WI -
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