High resolution separation of recombinant monoclonal antibodies by size-exclusion ultra-high performance liquid chromatography (SE-UHPLC)

Journal of Pharmaceutical and Biomedical Analysis
Renee YangYonghua Taylor Zhang

Abstract

Size-exclusion chromatography (SEC) is an important mode of separation used in monoclonal antibody (mAb) characterization and quality control. SEC separates mAbs into three major species: high molecular weight species, main peak (predominantly monomer), and low molecular weight species. However, mAb SEC separations have low resolution between the different sized species, and the analysis is slow with low sample throughput. The introduction of size-exclusion ultra-high performance liquid chromatography (SE-UHPLC) columns offers a new opportunity to improve both the resolution and throughput of SEC analysis. This study demonstrates that SE-UHPLC columns deliver better resolution of size variants in a shorter period of time than conventional SEC columns. For example, an SE-UHPLC column 300-mm in length produced separation of mAb Fab/c fragments in less than 10min, in comparison to a conventional SEC column output, where these fragments co-elute with the main peak. Furthermore, we observed that high back pressure does not generate HMWS under optimized mobile phase conditions for mAbs. The platform SE-UHPLC method has been demonstrated to be suitable for the analysis of multiple mAbs, with greatly improved sample throughput and peak...Continue Reading

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Citations

May 9, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Balázs BobályJenő Fekete
Jun 22, 2016·Journal of Pharmaceutical and Biomedical Analysis·Maria Kristina ParrHouda Montacir
Jul 28, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Tao ChenKelly Zhang
Jun 25, 2018·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Alexandre GoyonDavy Guillarme
Jul 10, 2017·Journal of Pharmaceutical and Biomedical Analysis·Balázs BobálySzabolcs Fekete
Dec 20, 2016·Journal of Pharmaceutical and Biomedical Analysis·Alexandre GoyonSzabolcs Fekete
Apr 13, 2021·Biotechnology and Bioengineering·Tingwei RenZheng Jian Li

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