KRIBIOLISA™ E.coli Host Cell Protein ELISA

Introduction:

A variety of proteins and pDNA which are used as therapeutic agents in humans and animals are produced through recombinant expression in E.coli. The manufacturing and purification process of these products tends to leave the potential for contamination by Host Cell Proteins (HCPs) from E.coli 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 generic kit is intended in determining the presence of E.coli Host Cell Proteins contamination in various products that are manufactured through recombinant expression in E.coli. The kit has been validated successfully for testing of final and in process product HCPs in variety of products regardless of growth and purification process.

Principle:

This assay is based on the Sandwich ELISA procedure. Samples containing E-coli HCPs are reacted with antiE.coli:HRP antibody simultaneously in the microtiter wells already coated with affinity purified capture anti-E.coli antibody. This immunological reaction results in formation of a sandwich complex of solid phase antibody-HCPenzyme labeled antibody. The wells are washed to remove any unbound reactants as per the wash procedure (see Assay procedure section mentioned below). The substrate 3,3’,5,5’ Tetramethyl 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 E.coli HCPs present.