ELISA Determines ChABC Distribution and Content in Tissues and Bodily Fluids
Chondroitinase ABC (ChABC) is a bacterial enzyme that degrades sugar chains attached to the core protein of chondroitin sulfate proteoglycans (CSPGs). CSPGs are involved in a variety of biological processes in higher organisms, including humans. ChABC has a well-known potential for therapeutic application and extensive testing is underway. Therefore, the ELISA has an important application to determine ChABC distribution and content in tissues and bodily fluids. A panel of monoclonal antibodies was produced that specifically bind ChABC. Additional screening identified a pair of complimentary monoclonal antibodies, a capture antibody and detection antibody, and a Sandwich ELISA was established. A working protocol for the Sandwich ELISA was established and validated. Extensive testing documents the accuracy, precision, selectivity, sensitivity and reproducibility of the ChABC ELISA methodology for quantifying and detecting ChABC in buffers and biological samples. The monoclonal anti-ChABC antibodies bind ChABC from prominent vendors.
In addition, both antibodies are effective in Western immunoblotting and immunohistochemistry. Detailed methods and assay specifications are available upon request. Presently, there is no commercial source of monoclonal antibody to ChABC. Efforts to quantify ChABC rely on assay of its enzymatic activity. This approach is complicated and lacks specificity when applied to complex biological samples and fails to detect inactive ChABC protein residuals. No quantitative immunoassays are available commercially for ChABC. We are seeking licensing partners to commercialize the ChABC antibodies and Sandwich ELISA. License of the hybridomas or purified antibodies will be considered.
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