A general Fc engineering platform for the next generation of antibody therapeutics.
A general Fc engineering platform for the next generation of antibody therapeutics.
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用于下一代抗体疗法的通用 Fc 工程平台
DOI：
10.7150/thno.51299
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发表时间：
2021
期刊：
影响因子：
12.4
通讯作者：
Zhang H
中科院分区：
文献类型：
--
作者：
Chen D;Zhao Y;Li M;Shang H;Li N;Li F;Wang W;Wang Y;Jin R;Liu S;Li X;Gao S;Tian Y;Li R;Li H;Zhang Y;Du M;Cao Y;Zhang Y;Li X;Huang Y;Hu LA;Li F;Zhang H
原理：Fc工程已成为抗体药物开发的焦点。目前的突变和计算机蛋白质设计方法受到通量有限和成本高的限制，而高通量噬菌体展示和酵母展示技术不适合筛选糖基化Fc变体。在这里，我们开发了一个基于哺乳动物细胞展示的Fc工程平台。研究方法：通过使用哺乳动物细胞展示和下一代测序，我们筛选了数百万种Fc变体，以优化对Fc γ RIIIa或Fc γ RIIb的亲和力和特异性。将鉴定的与Fc γ RIIIa结合改善的Fc变体替换为曲妥珠单抗和利妥昔单抗，并在基于PBMC的试验中检查抗体的效应子功能。另一方面，将鉴定的具有选择性增强的Fc γ RIIb结合的Fc变体应用于CD40激动剂抗体，并在不同的细胞测定中测量抗体的活性。还通过Fc γ R/CD40人源化小鼠中的OVA特异性CD8 + T细胞应答模型评价了CD40抗体的免疫刺激活性。结果如下：使用这种方法，我们筛选了数百万个Fc变体，并成功鉴定了几种具有增强Fc γ RIIIa或Fc γ RIIb结合的新型Fc变体。这些鉴定的Fc变体在基于PBMC的测定中显示抗体依赖性细胞毒性的显著增加。还鉴定了具有选择性增强的Fc γ RIIb结合的新型变体。用这些Fc变体取代的CD40激动剂抗体在体外和体内模型中显示出比亲本抗体更有效的活性。结论：这种方法将Fc变体筛选的通量从数千增加到数百万量级，使得能够筛选含有多个突变的变体，并且可以与糖工程技术整合，代表了Fc工程的理想平台。最初的努力证明了该平台的能力，并且新的Fc变体可以被替换成几乎任何抗体，用于下一代抗体治疗。
Rationale: Fc engineering has become the focus of antibody drug development. The current mutagenesis and in silico protein design methods are confined by the limited throughput and high cost, while the high-throughput phage display and yeast display technologies are not suitable for screening glycosylated Fc variants. Here we developed a mammalian cell display-based Fc engineering platform. Methods: By using mammalian cell display and next generation sequencing, we screened millions of Fc variants for optimized affinity and specificity for FcγRIIIa or FcγRIIb. The identified Fc variants with improved binding to FcγRIIIa were substituted into trastuzumab and rituximab and the effector function of antibodies were examined in the PBMC-based assay. On the other hand, the identified Fc variants with selectively enhanced FcγRIIb binding were applied to CD40 agonist antibody and the activities of the antibodies were measured on different cell assays. The immunostimulatory activity of CD40 antibodies was also evaluated by OVA-specific CD8+ T cell response model in FcγR/CD40-humanized mice. Results: Using this approach, we screened millions of Fc variant and successfully identified several novel Fc variants with enhanced FcγRIIIa or FcγRIIb binding. These identified Fc variants displayed a dramatic increase in antibody-dependent cellular cytotoxicity in PBMC-based assay. Novel variants with selectively enhanced FcγRIIb binding were also identified. CD40 agonist antibodies substituted with these Fc variants displayed activity more potent than the parental antibody in the in vitro and in vivo models. Conclusions: This approach increased the throughput of Fc variant screening from thousands to millions magnitude, enabled screening variants containing multiple mutations and could be integrated with glycoengineering technology, represents an ideal platform for Fc engineering. The initial efforts demonstrated the capability of the platform and the novel Fc variants could be substituted into nearly any antibody for the next generation of antibody therapeutics.
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影响因子：
8
作者：
Giles, Amber J.;Hao, Shuyu;Park, Deric M.
通讯作者：
Park, Deric M.
影响因子：
5.3
作者：
Kaplon, Helene;Muralidharan, Mrinalini;Reichert, Janice M.
通讯作者：
Reichert, Janice M.
DOI：
10.1126/science.1198443
发表时间：
2011-03-25
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Beatty GL;Chiorean EG;Fishman MP;Saboury B;Teitelbaum UR;Sun W;Huhn RD;Song W;Li D;Sharp LL;Torigian DA;O'Dwyer PJ;Vonderheide RH
通讯作者：
Vonderheide RH
影响因子：
7.2
作者：
Bajor DL;Mick R;Riese MJ;Huang AC;Sullivan B;Richman LP;Torigian DA;George SM;Stelekati E;Chen F;Melenhorst JJ;Lacey SF;Xu X;Wherry EJ;Gangadhar TC;Amaravadi RK;Schuchter LM;Vonderheide RH
通讯作者：
Vonderheide RH
影响因子：
4.8
作者：
Dall'Acqua, William F.;Kiener, Peter A.;Wu, Herren
通讯作者：
Wu, Herren