Massively parallel de novo protein design for targeted therapeutics.
Massively parallel de novo protein design for targeted therapeutics.
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DOI：
10.1038/nature23912
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发表时间：
2017-10-05
期刊：
影响因子：
64.8
通讯作者：
Baker D
中科院分区：
文献类型：
--
作者：
Chevalier A;Silva DA;Rocklin GJ;Hicks DR;Vergara R;Murapa P;Bernard SM;Zhang L;Lam KH;Yao G;Bahl CD;Miyashita SI;Goreshnik I;Fuller JT;Koday MT;Jenkins CM;Colvin T;Carter L;Bohn A;Bryan CM;Fernández-Velasco DA;Stewart L;Dong M;Huang X;Jin R;Wilson IA;Fuller DH;Baker D
从头蛋白质设计有望创造出具有定制形状以结合治疗靶点的小的稳定蛋白质。我们描述了一种用于设计、制造和筛选微型蛋白结合剂的大规模并行方法，该方法集成了大规模计算设计、寡核苷酸合成、酵母展示筛选和下一代测序。我们设计并测试了22，660个37-43个残基的迷你蛋白，其靶向流感血凝素和肉毒杆菌神经毒素B，沿着6，286个控制序列以探测对折叠和结合的贡献，并鉴定了2，618个高亲和力结合剂。结合和非结合的设计集，这是两个数量级大于任何以前的调查，使计算模型的评估和改进的比较。结合剂设计的子集的生物物理表征表明，它们非常稳定，并且与抗体不同，在暴露于高温后不会失去活性。该设计引起很少或没有免疫应答，并且即使在广泛重复给药后也提供针对流感的有效预防和治疗保护。
De novo protein design holds promise for creating small stable proteins with shapes customized to bind therapeutic targets. We describe a massively parallel approach for designing, manufacturing and screening mini-protein binders, integrating large-scale computational design, oligonucleotide synthesis, yeast display screening and next-generation sequencing. We designed and tested 22,660 mini-proteins of 37–43 residues that target influenza haemagglutinin and botulinum neurotoxin B, along with 6,286 control sequences to probe contributions to folding and binding, and identified 2,618 high-affinity binders. Comparison of the binding and non-binding design sets, which are two orders of magnitude larger than any previously investigated, enabled the evaluation and improvement of the computational model. Biophysical characterization of a subset of the binder designs showed that they are extremely stable and, unlike antibodies, do not lose activity after exposure to high temperatures. The designs elicit little or no immune response and provide potent prophylactic and therapeutic protection against influenza, even after extensive repeated dosing.
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影响因子：
64.8
作者：
Bhardwaj, Gaurav;Mulligan, Vikram Khipple;Bahl, Christopher D.;Gilmore, Jason M.;Harvey, Peta J.;Cheneval, Olivier;Buchko, Garry W.;Pulavarti, Surya V. S. R. K.;Kaas, Quentin;Eletsky, Alexander;Huang, Po-Ssu;Johnsen, William A.;Greisen, Per Jr;Rocklin, Gabriel J.;Song, Yifan;Linsky, Thomas W.;Watkins, Andrew;Rettie, Stephen A.;Xu, Xianzhong;Carter, Lauren P.;Bonneau, Richard;Olson, James M.;Coutsias, Evangelos;Correnti, Colin E.;Szyperski, Thomas;Craik, David J.;Baker, David
通讯作者：
Baker, David
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
DOI：
10.1073/pnas.1321126111
发表时间：
2014-06-10
影响因子：
11.1
作者：
King, Chris;Garza, Esteban N.;Baker, David
通讯作者：
Baker, David
影响因子：
4.4
作者：
JORGENSEN, WL;CHANDRASEKHAR, J;KLEIN, ML
通讯作者：
KLEIN, ML
影响因子：
4.4
作者：
ESSMANN, U;PERERA, L;PEDERSEN, LG
通讯作者：
PEDERSEN, LG