The Pichia pastoris expression system is a highly efficient eukaryotic expression system with post-translational processing modifications, which is commonly used for the stable expression of functional biological products such as recombinant proteins. Products using the Pichia pastoris expression system will inevitably introduce host cell protein (HCP) impurities during the production and purification process, which can reduce the efficacy of the product and lead to adverse toxicity or immune reactions, so it is necessary to ensure that the residual level of HCP in the final product of the biologic product is as low as possible. ELISA is a simple to use, sensitive and objective test for HCPs. This kit uses the principle of double-antibody sandwich enzyme-linked immunosorbent assay (sandwich ELISA) to detect the residual Pichia pastoris HCP in samples. First, add the Pichia pastoris HCP standard (36720-B) and test sample to the Anti-Pichia pastoris HCP coated microtiter strips (36720-A), then add the diluted biotin labeled Anti-Pichia pastoris HCP antibody (36720-C), and finally add Streptavidin-HRP (36720-D) to form an antibody + antigen + antibody-Biotin + SA-HRP complex. Subsequently, add TMB substrate (36720-H) into the complex to observe color reaction after washing the complex. TMB was converted from colorless to blue and finally to yellow in the presence of termination solution (36720-I) catalyzed by HRPase. Shades of yellow were positively correlated with the amount of Pichia pastoris HCP detected in the sample. This kit can be used for the optimization of the purification process of biologics, the control of impurities in intermediate processes, and the release testing of final products. The detection range of this kit is 3.125~200 ng/mL; the lower detection limit is 0.5 ng/mL. Features Highly sensitive: Detecting as little as 0.5 ng/mL of Pichia pastoris HCP residuals in in-process and final biological samples. Ensures accuracy: The Pichia pastoris HCP residuals can be accurately detected with a recovery of 75%-125%. Specificity: Specific detection of Pichia pastoris HCP without interference from other exogenous proteins. Stable: The difference between lot-to-lot is low, the kit property is not impacted under 37℃ for 7 days. Easy signal collection: Using Biotin-Streptavidin system, the signal can be stably enlarged. Application Residual Pichia pastoris HCP test in biological products Specification Sensitivity 0.5 ng/mL (range 3.125~200 ng/mL) Assay Time
