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- W2350962061 abstract "Objective To establish the method and standard for quality control of recombinant human monoclonal antibody against rabies (rhRMcAb). Methods The rhRMcAb was analyzed for neutralizing activity by mouse neutralization test(MNT)and rapid fluorescent focus inhibition test(RFFIT), for purity by reduced and non-reduced SDS-PAGE and for peptide map by reduction alkylation and trypsin digestion followed by reverse phase HPLC. The amino acids at N-terminus was sequenced by amino acid sequencer after the pyroglutamate blockage at N-terminus was removed by pyroglutamate aminopeptidase. The affinity constant of rhRM-cAb with rabies virus glycoprotein was determined by surface plasmon resonance(SPR)method. Other quality indexes of rhRMcAb were determined according to the requirements in Chinese Pharmacopeia (2005 edition), based on which a standard for quality control was established. Results The activities of six batches of rhRMcAb determined by MNT and RFFIT were basically in agreement. The reduced SDS-PAGE purities of three batches of rhRMcAb bulks were more than 99. 0%. Except major antibody bands, some minor bands with both high and low relative molecular masses were also observed on the non-reduced SDS-PAGE profile of rhRMcAb. The peptide maps of three batches of bulks were consistent with that of in-house reference. The amino acid sequence at N-terminus was identical to the theoretical value. The affinity constant of rhRMcAb with rabies virus glycoprotein was 1. 86 × 108 / M. All the other quality indexes of rhRMcAb met the requirements in Chinese Pharmacopeia(2005 edition). In the established standard for quality control, the neutralizing activity of rhRMcAb was not less than 500 IU / ml, while both SDS-PAGE and HPLC purities were not less than 98. 0%. Conclusion The method and standard for quality control of rhRMcAb were established." @default.
- W2350962061 created "2016-06-24" @default.
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- W2350962061 date "2010-01-01" @default.
- W2350962061 modified "2023-09-23" @default.
- W2350962061 title "Establishment of Method and Standard for Quality Control of Recombinant Human Monoclonal Antibody against Rabies" @default.
- W2350962061 hasPublicationYear "2010" @default.
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