KRIBIOLISA™ Pichia Pastoris HCP ELISA

Introduction:

A variety of proteins and pDNA which are used as therapeutic agents in humans and animals are produced through recombinant expression in P.pastoris. The manufacturing and purification process of these products tends to leave the potential for contamination by Host Cell Proteins (HCPs) from P.pastoris which may result in adverse toxic or immunological reactions that ultimately affect the efficacy of the therapeutic agent. The simple, objective and semi quantitative ELISA is a highly sensitive method that aids in purification process development, process control, quality control and product release testing optimally.

Intended Use:

This KRIBIOLISA™ Pichia Pastoris HCP ELISA is a generic kit intended in determining the presence of P.pastoris Host Cell Proteins contamination in various biotechnology products that are manufactured through recombinant expression in P. pastoris. The kit has been validated successfully for testing of in process and final product HCPs in variety of products regardless of growth and purification process.

Principle:

This assay is based on the Sandwich ELISA procedure. Samples containing P.pastoris HCPs are reacted with already coated affinity purified capture anti-P.pastoris antibody and bind to them. Plates are washed with wash buffer to remove unbound reactants. Anti P.pastoris-HRP conjugate is added which results in the formation of a sandwich complex of solid phase antibody-HCP-enzyme labeled antibody. The wells are washed to remove any unbound reactants as per the wash procedure. The substrate 3, 3’,5, 5’ Tetra Methyl Benzidine is then reacted. The amount of hydrolyzed substrate is read on a microtiter plate reader and it is directly proportional to the concentration of P. pastoris HCPs present.