Modeling the impact of amino acid substitution in a monoclonal antibody on cation exchange chromatography
| Autor: | Jürgen Hubbuch, Michelle Ahlers-Hesse, Rudger Hess, Joschka Bauer, David Saleh, Simon Kluters, Federico Rischawy, Nicole Beckert, Gang Wang, Markus Schönberger, Michaela Blech, Joey Studts |
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| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
0301 basic medicine Models Molecular medicine.drug_class Lysine Ion chromatography Bioengineering Monoclonal antibody Immunoglobulin light chain 01 natural sciences Applied Microbiology and Biotechnology 03 medical and health sciences 010608 biotechnology medicine Homology modeling chemistry.chemical_classification Downstream processing Elution Antibodies Monoclonal Chromatography Ion Exchange Complementarity Determining Regions Amino acid 030104 developmental biology chemistry Biochemistry Amino Acid Substitution Immunoglobulin G Immunoglobulin Light Chains Immunoglobulin Heavy Chains Biotechnology |
| Zdroj: | Biotechnology and bioengineeringREFERENCES. 118(8) |
| ISSN: | 1097-0290 |
| Popis: | A vital part of biopharmaceutical research is decision making around which lead candidate should be progressed in early-phase development. When multiple antibody candidates show similar biological activity, developability aspects are taken into account to ease the challenges of manufacturing the potential drug candidate. While current strategies for developability assessment mainly focus on drug product stability, only limited information is available on how antibody candidates with minimal differences in their primary structure behave during downstream processing. With increasing time-to-market pressure and an abundance of monoclonal antibodies (mAbs) in development pipelines, developability assessments should also consider the ability of mAbs to integrate into the downstream platform. This study investigates the influence of amino acid substitutions in the complementarity-determining region (CDR) of a full-length IgG1 mAb on the elution behavior in preparative cation exchange chromatography. Single amino acid substitutions within the investigated mAb resulted in an additional positive charge in the light chain (L) and heavy chain (H) CDR, respectively. The mAb variants showed an increased retention volume in linear gradient elution compared with the wild-type antibody. Furthermore, the substitution of tryptophan with lysine in the H-CDR3 increased charge heterogeneity of the product. A multiscale in silico analysis, consisting of homology modeling, protein surface analysis, and mechanistic chromatography modeling increased understanding of the adsorption mechanism. The results reveal the potential effects of lead optimization during antibody drug discovery on downstream processing. |
| Databáze: | OpenAIRE |
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