A Rapid Analysis Method for Monoclonal Antibody N-Glycoforms

Abstract

The invention provides a rapid analysis method for monoclonal antibody N sugar glycoform. The analysis method comprises the following steps: after centrifuging a to-be-tested cell culture, taking thesupernatant, diluting, adding an IdeS enzyme and a reducing agent to perform enzymatic hydrolysis reaction and reduction reaction, centrifuging, and taking the supernatant; detecting an antibody digestion and a reducing subunit solution by using capillary electrophoresis mass spectrometry to obtain the intensity and molecular weight of different glycoform molecules Ai in the antibody digestion andthe reduction subunit solution; determining the glycoforms corresponding to the molecular weights of the different glycoform molecules Ai by querying a database containing molecular weights of various glycoform molecules, and taking the ratio of the total strength of the subunits to which the intensity of each glycoform molecule Ai belongs as its glycoform ratio. Compared with the traditional polysaccharide labeling method, the rapid analysis method for the monoclonal antibody N sugar glycoform has advantages of rapid, high efficiency and sensitivity, provides strong support for the development of cell strains in the biopharmaceutical industry, and will greatly assist in speeding up the development progress of cell strain screening.

Description

technical field [0001] The invention relates to the technical field of chemical analysis and detection, in particular to a rapid analysis method for N-glycoforms of monoclonal antibodies. Background technique [0002] In the pharmaceutical industry, biomedical technology is receiving increasing attention. The use of monoclonal antibody has a history of more than 100 years. Due to its low toxicity and side effects, high specific efficacy, long half-life and easy large-scale production, monoclonal antibody has become a very mature drug in the field of biopharmaceuticals. It is also an area that many pharmaceutical companies are vying to set foot in. At present, most monoclonal antibody products are expressed by mammalian cells. A large part of the reason is that mammalian cells can achieve complex glycosylation modifications on monoclonal antibody protein molecules, and glycosylation modifications can increase protein stability and solubility. , in order to achieve a longer ...

AI Technical Summary

Problems solved by technology

[0006] ②The process of sugar cutting and derivatization reagent labeling is time-consuming and laborious;

[0008] Therefore, using the traditional 2-AA or 2-AB labeling method, a single sample analysis test takes a long time, resulting in a small amount of detection and analysis samples per unit time, low efficiency, low detection throughput and low degree of automation

It takes about 3 days from getting the sample to getting the result, which causes a great waste of time for the screening of monoclonal antibody cell lines

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