Structural basis of a novel heterodimeric Fc for bispecific antibody production.
Structural basis of a novel heterodimeric Fc for bispecific antibody production.
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用于双特异性抗体生产的新型异二聚体 Fc 的结构基础
DOI：
10.18632/oncotarget.17558
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发表时间：
2017-08-01
期刊：
影响因子：
--
通讯作者：
Zhou A
中科院分区：
文献类型：
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作者：
Wei H;Cai H;Jin Y;Wang P;Zhang Q;Lin Y;Wang W;Cheng J;Zeng N;Xu T;Zhou A
双特异性抗体为联合临床治疗提供了一种有效工具。在此，我们通过结合“嵌合孔穴”和静电导向策略，设计了一种用于生产双特异性抗体的异源二聚体Fc片段。其中，IgG CH3结构域界面上一个较大的疏水残基苯丙氨酸405（Phe405）突变为带电荷的残基赖氨酸（Lys），相应CH3结构域中的赖氨酸409（Lys409）突变为丙氨酸（Ala）。此处解析得到的该Fc异源二聚体的晶体结构，分辨率为2.7埃，揭示了这两个突变如何形成互补的结合界面，并解释了为何F405K突变能在蛋白质表达过程中有效抑制Fc同源二聚体的形成。利用这种异源二聚体Fc构建出一种源自曲妥珠单抗和帕妥珠单抗的抗HER2双特异性抗体。在体外和体内抑制癌细胞增殖方面，该双特异性抗体展现出与曲妥珠单抗和帕妥珠单抗联合使用相当或更优的效果。总体而言，本研究表明，此处设计的异源二聚体Fc为生成用于癌症治疗的活性双特异性抗体提供了一个高效平台。
Bispecific antibodies provide an efficient tool for combinational clinical therapy. Here we have engineered a heterodimeric Fc for bispecific antibodies production by combining the knob-into-hole and electrostatic steering strategies where a bulky hydrophobic residue Phe405 of the IgG CH3 interface is mutated to a charged residue Lys and Lys409 of the corresponding CH3 domain is mutated to Ala. The crystal structure of this Fc heterodimer solved here at 2.7Å resolution revealed how these two mutations resulted a complementary binding interface and explained why F405K mutation could effectively inhibit Fc homodimer formation during protein expression. An anti-HER2 bispecific antibody derived from trastuzumab and pertuzumab was generated by this heterodimeric Fc. It showed comparable or improved efficacy than the combination of trastuzumab and pertuzumab in inhibiting proliferation of cancer cells in vitro and in vivo. Overall this study shows that the heterodimeric Fc engineered here provides an efficient platform for generating active bispecific antibody for cancer treatment.