Method for effectively removing host protein in monoclonal antibody downstream purification process

Abstract

The invention relates to a method for effectively removing a host protein in a monoclonal antibody downstream purification process. The method comprises the following steps: (1) performing deep filtration on cell culture fluid, and collecting the filtrate; (2) performing Protein A affinity chromatography on the filtrate obtained in the step (1); (3) performing virus inactivation on the sample obtained in the step (2), performing deep filtering on the sample subjected to virus inactivation, thereby effectively removing the host protein in the sample containing the monoclonal antibody. According to the method disclosed by the invention, the deep filtering after virus inactivation is adopted, the deep filtering process after Protein A affinity chromatography and virus inactivation is optimized, the content of HCP in the culture fluid supernatant collected by deep filtering can be reduced to 2.3ppm by virtue of two purification steps, namely the Protein A affinity chromatography and deep filtering after virus inactivation, and the content of the HCP can be reduced to 0.4ppm by virtue of further anion and cation exchange chromatography.

Description

technical field [0001] The present invention relates to the purification technology of monoclonal antibody, and more specifically relates to the method for effectively removing host protein in the downstream purification process of monoclonal antibody. Background technique [0002] At present, the downstream process platform of monoclonal antibodies generally mainly includes fermentation broth clarification, capture steps, purification steps, virus inactivation and filtration, and ultrafiltration concentration. [0003] With the development of large-scale CHO cell culture technology, the protein expression level of cells has been greatly improved, but with the large amount of exogenous nutrients added and the cell density increased by orders of magnitude during the culture process, the components in cell culture products It's getting more and more complicated. Host Cell Protein (HCP) is one of the complex pollutants. The complexity and heterogeneity of HCP depends on the ex...

AI Technical Summary

Problems solved by technology

[0007] In the traditional monoclonal antibody process platform, several downstream purification processes, such as depth filtration, Protein A affinity chromatography, one-step or two-step purification chromatography, etc., all need to effectively remove HCP, so as to gradually reduce the HCP content to a manageable level. Within the accepted standard, but the content of HCP is difficult to be reduced to 10ppm, especially below 1ppm

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