Comparison of three types of residual Chinese hamster ovary cell protein detection kits / 中国药学杂志

ABSTRACT

OBJECTIVE: To compare the detection ability of three types of residual Chinese hamster ovary(CHO) host cell protein (HCP) ELISA kits (Cat No. A, B, C) from one manufacturer. METHODS: Residual CHO-HCP in the bulks of therapeutic monoclonal antibody (mcAb) produced with different strains of CHO cells, extracted proteins from CHO-K1 cultural supernatant and standards from different kits were determined with three types of kits to evaluate the CHO-HCP detection ability of different kits. Furthermore, the capture antibody coated microplates and enzyme conjugated detection antibody of three kits were re-assembled to detect standards and extracted supernatant proteins to analyze the possible reasons of differentiated detection ability. RESULTS: For the detection of mcAb samples, the relative recoveries of kit A and B to kit C were 1.89% and 15.34%, respectively; for the detection of extracted proteins from CHO-K1 cultural supernatant, the recoveries of kit A, B, and C were 4.05%, 21.52%, and 35.98%, respectively; for the detection of self-standards, the recoveries of three kits were all within the range of (100±15)%. For the detection of standards from other kits, there was a great discrepancy among the three kits. For kit A detection, the recoveries of standard B and C were 31.28% and 7.99%; for kit B detection, the recoveries of standard A and C were 55.76% and 25.08%; for kit C detection, the recoveries of standard A and B were 266.65% and 73.31%, respectively. By analyzing the results from reassembled capture and secondary antibodies, it was inferred that the detection ability of the capture antibodies of the three kits was sequenced as A>C>B, while the detection ability of enzyme conjugated secondary antibodies was sequenced as B>C>A. CONCLUSION: The detection abilities of three CHO-HCP ELISA kits from the investigated manufacturer were determined as C>B>A.