Enhancing Antibody Fc Heterodimer Formation through Electrostatic Steering Effects APPLICATIONS TO BISPECIFIC MOLECULES AND MONOVALENT IgG
Enhancing Antibody Fc Heterodimer Formation through Electrostatic Steering Effects APPLICATIONS TO BISPECIFIC MOLECULES AND MONOVALENT IgG
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DOI：
10.1074/jbc.m110.117382
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发表时间：
2010-06-18
影响因子：
4.8
通讯作者：
Yan, Wei
中科院分区：
文献类型：
--
作者：
Gunasekaran, Kannan;Pentony, Martin;Yan, Wei
天然存在的IgG抗体是二价和单特异性的。对两种不同抗原具有结合特异性的双特异性抗体可以使用重组技术产生，并且预计具有广泛的临床应用。然而，多个轻链和重链的共表达通常导致污染物并造成纯化挑战。在这项工作中，我们用选定的突变修饰了抗体Fc区的CH3结构域界面，使得工程化的Fc蛋白优先形成异二聚体。这些新的突变产生改变的电荷极性跨越Fc二聚体界面，使得静电匹配的Fc链的共表达支持有利的吸引相互作用，从而促进所需的Fc异二聚体形成，而不利的排斥电荷相互作用抑制不需要的Fc同二聚体形成。这种新的Fc异二聚体形式用于产生具有最少同源二聚体污染物的双特异性单链抗体融合物和单价IgG。本文提出的策略证明了稳健生产新型基于Fc的异二聚体分子的可行性，并因此拓宽了用于治疗应用的双特异性分子的范围。
Naturally occurring IgG antibodies are bivalent and monospecific. Bispecific antibodies having binding specificities for two different antigens can be produced using recombinant technologies and are projected to have broad clinical applications. However, co-expression of multiple light and heavy chains often leads to contaminants and pose purification challenges. In this work, we have modified the CH3 domain interface of the antibody Fc region with selected mutations so that the engineered Fc proteins preferentially form heterodimers. These novel mutations create altered charge polarity across the Fc dimer interface such that coexpression of electrostatically matched Fc chains support favorable attractive interactions thereby promoting desired Fc heterodimer formation, whereas unfavorable repulsive charge interactions suppress unwanted Fc homodimer formation. This new Fc heterodimer format was used to produce bispecific single chain antibody fusions and monovalent IgGs with minimal homodimer contaminants. The strategy proposed here demonstrates the feasibility of robust production of novel Fc-based heterodimeric molecules and hence broadens the scope of bispecific molecules for therapeutic applications.