文章基本信息

标题：Next generation Fc scaffold for multispecific antibodies
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作者：Bram Estes ; Athena Sudom ; Danyang Gong 等
期刊名称：iScience
印刷版ISSN：2589-0042
出版年度：2021
卷号：24
期号：12
页码：1-21
DOI：10.1016/j.isci.2021.103447
语种：English
出版社：Elsevier
摘要：SummaryBispecific antibodies (Bispecifics) demonstrate exceptional clinical potential to address some of the most complex diseases. However, Bispecific production in a single cell often requires the correct pairing of multiple polypeptide chains for desired assembly. This is a considerable hurdle that hinders the development of many immunoglobulin G (IgG)-like bispecific formats. Our approach focuses on the rational engineering of charged residues to facilitate the chain pairing of distinct heavy chains (HC). Here, we deploy structure-guided protein design to engineer charge pair mutations (CPMs) placed in the CH3-CH3′ interface of the fragment crystallizable (Fc) region of an antibody (Ab) to correctly steer heavy chain pairing. When used in combination with our stable effector functionless 2 (SEFL2.2) technology, we observed high pairing efficiency without significant losses in expression yields. Furthermore, we investigate the relationship between CPMs and the sequence diversity in the parental antibodies, proposing a rational strategy to deploy these engineering technologies.Graphical abstractDisplay OmittedHighlights•Crystal structures unveil molecular basis of SEFL2.2 and CPM technologies•Next gen structure-guided design of CPMs to steer HC-HC pairing•Top CPMs show high pairing efficiency and optimal expression and stability•Balancing CPM charge distribution minimizes impact of sequence diversityBiochemistry; Bioengineering; Biomolecular engineering; Structural biology