Chen Yu-Cheng, Zhong Xue-Zhao, Shi Ce, Chen Ran, Sponchioni Mattia, Yao Shan-Jing, Lin Dong-Qiang
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.
Shanghai Engineering Research Center of Anti-tumor Biological Drugs, Shanghai Henlius Biotech, Inc., Shanghai, China.
J Chromatogr A. 2025 Jan 25;1741:465602. doi: 10.1016/j.chroma.2024.465602. Epub 2024 Dec 16.
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 R 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 %).
