Study On Related Methods Of Quality Control Of Monoclonal Antibody Drugs

Monoclonal antibodies(mAbs),have been effectively utilized in the management of several malignancies,in transplant rejection,in autoimmune and inflammatory diseases,and in a range of further indications.Monoclonal antibody drugs occupy an increasing share of the biopharmaceutical market,and more and more companies are adding to the production of monoclonal antibodies,At the same time,in order to ensure the safety and effectiveness of monoclonal antibody drug products and to promote orderly research and development and competition in this field,relevant regulatory authorities continue to put forward new requirements for quality control of monoclonal antibody drugs.This paper uses a variety of analytical methods to characterize the purity,extinction coefficient and activity separation of monoclonal antibody drugs,and provides some reference for the quality control and detection methods of monoclonal antibody drugs.The specific research contents are as follows:1.Monoclonal drug products may form size variants such as precursors,certain degradation products during production and/or storage.The phenomenon of the variant will cause certain damage to the monoclonal antibody drug,such as lowering the concentration of the protein monomer in the drug,thereby reducing the activity of the protein drug.Therefore,the purity characterization of the monoclonal antibody and the detection of size variants are particularly important.In this paper,the incubation temperature,time and pH value of the sample under non-reducing conditions under sodium dodecyl sulfate capillary electrophoresis were optimized.The purity and size variants of the monomer under the conditions of capillary electrophoresis reduction and non-reduction were combined with size exclusion chromatography using UV,differential,multi-angle static light scattering detector.The combined use of the three methods increases the accuracy of sample purity characterization.2.The extinction coefficient of the monoclonal antibody is not only one of the important physicochemical constants of the monoclonal antibody but also the key to the accuracy of the UV spectrophotometric value.In this thesis,the UV spectrophotometry,amino acid analyzer method and SEC-UV-RI-MALS method were used to calibrate the extinction coefficient of the antibody.The difference between the theoretical extinction coefficient and the measured extinction coefficient was compared.The setting of the drug extinction coefficient provides a reference.After calculating the difference value is 0.031%?5.814%,the measured extinction coefficient of the monoclonal antibody sample has little difference from the theoretical extinction coefficient.The mass concentration of the sample under ultraviolet spectrophotometry and the uncertainty of the method were also calculated.3.Commonly used methods for measuring activity such as cytotoxicity test,difference in cell proliferation in vitro,and enzyme-linked immunosorbent assay have problems such as poor reproducibility,low precision,complicated operation,and possible destruction of protein activity.In view of this situation,this paper selects the size exclusion chromatography method with moderate separation conditions,does not destroy the protein structure,and asymmetric flow field flow technology to separate and characterize the monoclonal antibody samples,optimize the key parameters separately,and determine the resolution and recovery rate.The optimal conditions have improved the repeatability and precision of the analytical methods,and provided new ideas for the subsequent characterization of antibody activity.It also opened a new perspective for the use of other detectors to achieve accurate quality control of monoclonal antibodies.