Quality control study of recombinant anti-amyloid-β monoclonal antibody
Objective:To study and establish a quality control method for the critical quality attributes of anti-amyloid-β monoclonal antibody(anti-Aβ mAb).Methods:Peptide fingerprints assay based on trypsin digestion and reversed-phase HPLC(RP-HPLC)was used for identification.The purities of monomers and polymers were analyzed by size-exclusion HPLC(SE-HPLC).Reducing/non-reducing capillary electrophoresis-sodium dodecyl sulfonate(CE-SDS)was used to analyze the purities of the size variants.The purities of the charge variants were analyzed by cationic chromatography(CEX-HPLC).The oligosaccharides were analyzed by HILIC-UPLC.The relative binding activity of Aβ antigen was determined by ELISA.All other indicators should comply with the current version of the Pharmacopoeia of the People's Republic of China and other relevant requirements.Results:The fingerprint of peptide achieved good identification.The relative percentage of SE-HPLC monomer was(99.40±0.00)％and the relative percentage of polymers was(0.55±0.00)％.The relative percentage of light and heavy chains in reduced CE-SDS was(98.11±0.08)％,the relative percentage of main peak was(96.57±0.15)％and the relative percentage of HHL peak was(1.91±0.10)％in non-reduced CE-SDS.The relative percentages of principal component,acid variant and basic variant were(75.08±0.06)％,(16.12±0.05)％and(8.80±0.05)％respectively in CEX-HPLC.The relative percentages of(G0F+G1F+G2F),Man5 and G0 were(91.56±0.04)％,(1.21±0.03)％and(2.60±0.01)％,respectively.The relative binding activity to Aβ antigen by ELISA was(101.40±0.53)％.Conclusion:According to the quality attribute of anti-Aβ monoclonal antibody,the quality control methods were established to ensure the safety,effectiveness and quality control of the product and provide a reference for the quality control methods and strategies of this kind of monoclonal antibody products.The study is of great significance to be a representative quality control reference for anti-Aβ mAbs.
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