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- 저자 : Séverine Fagète (검색결과 1 건)
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Célia De Temmerman,Pauline Bernard,Quentin Aeberli,Sarah Braito,Ghislaine Arib,Pierre-Alain Girod,Séverine Fagète 한국생물공학회 2023 Biotechnology and Bioprocess Engineering Vol.28 No.4
Bispecific antibodies (bsAb) represent the most promising class of therapeutics due to their multiple mode of action. Their development raised tremendous efforts with a special focus on the engineering of these complex molecules, especially when they adopt an asymmetrical format. Various strategies have been successful in improving heterodimerization through the introduction of point mutations to force the correct pairing of opposite chains using either steric, electrostatic steering effects or both. These great advances in protein engineering combined with the development of robust process purification methods to remove homodimers have contributed to the fast progress of bsAbs towards clinics. Nevertheless, there is still a lack of systematic and efficient production methods to fine tune the expression of correctly paired bsAbs. In this study, we have evaluated the effects of manipulating different molar ratios of three polypeptide chains constituting a model bsAb, co-transfected, expressed and assembled in a single Chinese Hamster Ovary cell line. Two parental clones showing different heterodimerization and productivity profiles were chosen. We demonstrated that addition of gene copies at specific ratios to these clones could bias towards the generation of sub-clones with either reduced proper assembly or tune expression for enhanced integral bsAb. Our results demonstrate that expression of bsAb can be influenced beyond the initial engineering stage.
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