DiPODS: A Reagent for Site-Specific Bioconjugation via the Irreversible Rebridging of Disulfide Linkages.
DiPODS: A Reagent for Site-Specific Bioconjugation via the Irreversible Rebridging of Disulfide Linkages.
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DOI：
10.1021/acs.bioconjchem.0c00590
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发表时间：
2020-12-16
影响因子：
4.7
通讯作者：
Price EW
中科院分区：
文献类型：
--
作者：
Khozeimeh Sarbisheh E;Dewaele-Le Roi G;Shannon WE;Tan S;Xu Y;Zeglis BM;Price EW
与硫醇的化学选择性反应长期以来一直有望用于抗体和抗体片段的位点特异性生物缀合。然而，带有单价马来酰亚胺的双功能探针-长期以来是基于硫醇的连接的“金标准”-受到两个内在问题的阻碍：马来酰亚胺-硫醇键的体内不稳定性和永久破坏二硫键以促进生物缀合的需要。在此，我们提出了合成，表征和验证的二PODS，一种新型的生物共轭试剂含有一对恶二唑基甲基砜部分能够不可逆地形成共价键与两个硫醇基团，同时重新桥接二硫键。该试剂是从市售的起始材料在8个步骤中合成的，在此期间遇到旋转异构体，并进行了实验和计算研究。DiPODS被设计为模块化的，因此可以通过悬垂的末端伯胺（DiPODS-PEG 4-NH 2）与任何有效负载缀合。随后，用荧光素缀合的DiPODS变体（DiPODS-PEG 4-FITC）修饰HER 2靶向Fab增强了试剂的位点特异性，说明了其重新桥接二硫键的能力，并产生了体外性质优于使用传统随机生物缀合技术产生的类似构建体的免疫缀合物上级。最终，我们认为这项工作对免疫缀合物的合成具有特别重要的意义，特别是确保货物与免疫球蛋白的连接是稳健的，不可逆的，并且在生物学和结构上是良性的。我们提出了合成，表征和验证的二PODS，一种新型的生物共轭试剂含有一对恶二唑基甲基砜部分，能够不可逆地形成共价键与两个硫醇基团，同时重新桥接二硫键。
Chemoselective reactions with thiols have long held promise for the site-specific bioconjugation of antibodies and antibody fragments. Yet bifunctional probes bearing monovalent maleimides — long the ‘gold standard’ for thiol-based ligations — are hampered by two intrinsic issues: the in vivo instability of the maleimide-thiol bond and the need to permanently disrupt disulfide linkages in order to facilitate bioconjugation. Herein, we present the synthesis, characterization, and validation of DiPODS, a novel bioconjugation reagent containing a pair of oxadiazolyl methyl sulfone moieties capable of irreversibly forming covalent bonds with two thiolate groups while simultaneously re-bridging disulfide linkages. The reagent was synthesized from commercially available starting materials in 8 steps, during which rotamers were encountered and investigated both experimentally and computationally. DiPODS is designed to be modular and can thus be conjugated to any payload through a pendant terminal primary amine (DiPODS–PEG4-NH2). Subsequently, the modification of a HER2-targeting Fab with a fluorescein-conjugated variant of DiPODS (DiPODS-PEG4-FITC) reinforced the site-specificity of the reagent, illustrated its ability to rebridge disulfide linkages, and produced an immunoconjugate with in vitro properties superior to those of an analogous construct created using traditional stochastic bioconjugation techniques. Ultimately, we believe that this work has particularly important implications for the synthesis of immunoconjugates, specifically for ensuring that the attachment of cargoes to immunoglobulins is robust, irreversible, and biologically and structurally benign. We present the synthesis, characterization, and validation of DiPODS, a novel bioconjugation reagent containing a pair of oxadiazolyl methyl sulfone moieties that is capable of irreversibly forming covalent bonds with two thiolate groups while simultaneously re-bridging disulfide linkages.
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影响因子：
3.1
作者：
Adumeau P;Sharma SK;Brent C;Zeglis BM
通讯作者：
Zeglis BM
影响因子：
4.7
作者：
Li X;Patterson JT;Sarkar M;Pedzisa L;Kodadek T;Roush WR;Rader C
通讯作者：
Rader C
影响因子：
4.9
作者：
Adumeau P;Vivier D;Sharma SK;Wang J;Zhang T;Chen A;Agnew BJ;Zeglis BM
通讯作者：
Zeglis BM
影响因子：
4.7
作者：
Bryden, Francesca;Maruani, Antoine;Boyle, Ross W.
通讯作者：
Boyle, Ross W.
影响因子：
4.7
作者：
Badescu, George;Bryant, Penny;Godwin, Antony
通讯作者：
Godwin, Antony