Mechanistic modeling of anti-Langmuirian to Langmuirian behavior of Fc-fusion proteins in cation exchange chromatography

Yu-Cheng Chen; Xue-Zhao Zhong; Ce Shi; Ran Chen; Mattia Sponchioni; Shan-Jing Yao; Dong-Qiang Lin

doi:10.1016/j.chroma.2024.465602

Mechanistic modeling of anti-Langmuirian to Langmuirian behavior of Fc-fusion proteins in cation exchange chromatography

J Chromatogr A. 2024 Dec 16:1741:465602. doi: 10.1016/j.chroma.2024.465602. Online ahead of print.

Authors

Yu-Cheng Chen¹, Xue-Zhao Zhong², Ce Shi², Ran Chen², Mattia Sponchioni³, Shan-Jing Yao⁴, Dong-Qiang Lin⁵

Affiliations

¹ Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang Key Laboratory of Smart Biomaterials, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310058, China; Department of Chemistry, Materials and Chemical Engineering, Politecnico di Milano, Via Mancinelli 7 20131 Milano, Italy.
² Shanghai Engineering Research Center of Anti-tumor Biological Drugs, Shanghai Henlius Biotech, Inc., Shanghai, China.
³ Department of Chemistry, Materials and Chemical Engineering, Politecnico di Milano, Via Mancinelli 7 20131 Milano, Italy.
⁴ Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang Key Laboratory of Smart Biomaterials, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310058, China.
⁵ Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang Key Laboratory of Smart Biomaterials, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310058, China. Electronic address: lindq@zju.edu.cn.

PMID: 39740612
DOI: 10.1016/j.chroma.2024.465602

Abstract

Development of a next-generation chromatographic model, capable of simultaneously meeting academic demands for thermodynamic consistency and industrial requirements in everyday project work, has become a focal point of research. In this study, anti-Langmuirian to Langmuirian (AL-L) elution behavior was observed in cation-exchange chromatographic separation of charge variants of industrial Fc-fusion proteins. To characterize this behavior, the multi-protein Mollerup activity model was integrated into the steric mass action (SMA) model, resulting in a new model named the generalized ion-exchange (nGIEX) isotherm for multi-protein systems. An R² exceeding 0.95 calibrated by three elution experiments indicates an effective description of the AL-L behavior (dynamic adsorption). Using isotherm sampling, the nGIEX model exhibited sigmoidal AL-L isotherms (static adsorption). Finally, the model's extrapolation capability was externally validated through process optimization, resulting in an optimal two-step elution condition and a yield improvement of the main variant from 25.9 % to 89.1 % within purity specifications (>70 %).

Keywords: Fc-fusion protein; anti-Langmuirian to Langmuirian behavior; ion-exchange chromatography; mechanistic model; process optimization; thermodynamic model.