General and Robust Chemoenzymatic Method for Glycan-Mediated Site-Specific Labeling and Conjugation of Antibodies: Facile Synthesis of Homogeneous Antibody-Drug Conjugates.
General and Robust Chemoenzymatic Method for Glycan-Mediated Site-Specific Labeling and Conjugation of Antibodies: Facile Synthesis of Homogeneous Antibody-Drug Conjugates.
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DOI：
10.1021/acschembio.1c00597
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发表时间：
2021-11-19
影响因子：
4
通讯作者：
Wang LX
中科院分区：
文献类型：
--
作者：
Zhang X;Ou C;Liu H;Prabhu SK;Li C;Yang Q;Wang LX
抗体的位点特异性标记和缀合对于基础研究和开发更有效的诊断和治疗方法是非常期望的。我们在这里报告了一个通用的和强大的化学酶的方法，允许一锅位点特异性功能化的抗体。设计、合成了一系列选择性修饰的二糖恶唑啉衍生物，并评价了其作为不同糖苷内切酶的供体底物用于抗体Fc聚糖重塑。我们发现，在测试的几种内切糖苷酶中，来自血清型M49的化脓链球菌的野生型内切糖苷酶（Endo-S2）在将携带位点选择性修饰的叠氮化物、生物素或荧光标签的官能化二糖转移至抗体而不水解所得的转糖基化产物方面表现出显著的活性。这一发现以及Endo-S2对重组抗体的优异Fc去糖基化活性，允许以一锅法直接标记和功能化抗体，而无需中间体和酶分离。不同数量的叠氮基的位点特异性引入使得能够通过无铜的应变促进的点击反应高效合成均质抗体-药物缀合物（ADC），其中药物与抗体的比率（DAR）精确控制在2至12的范围内。细胞活力测定显示，具有较高DAR的ADC比具有较低DAR的ADC更有效地杀死抗原过表达的细胞。这种新方法不仅有望应用于抗体-药物偶联，还有望应用于细胞标记、成像和诊断。
Site-specific labeling and conjugation of antibodies are highly desirable for fundamental research and for developing more efficient diagnostic and therapeutic methods. We report here a general and robust chemoenzymatic method that permits a one-pot site-specific functionalization of antibodies. A series of selectively modified disaccharide oxazoline derivatives were designed, synthesized, and evaluated as donor substrates of different endoglycosidases for antibody Fc glycan remodeling. We found that among several endoglycosidases tested, wild-type endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) exhibited remarkable activity in transferring the functionalized disaccharides carrying site-selectively modified azide, biotin, or fluorescent tags to antibodies without hydrolyzing the resulting transglycosylation products. This discovery, together with the excellent Fc deglycosylation activity of Endo-S2 on recombinant antibodies, allowed direct labeling and functionalization of antibodies in a one-pot manner without the need of intermediate and enzyme separation. The site-specific introduction of varied numbers of azide groups enabled a highly efficient synthesis of homogeneous antibody–drug conjugates (ADCs) with a precise control of the drug-to-antibody ratio (DAR) ranging from 2 to 12 via a copper-free strain-promoted click reaction. Cell viability assays showed that ADCs with higher DARs were more potent in killing antigen-overexpressed cells than the ADCs with lower DARs. This new method is expected to find applications not only for antibody–drug conjugation but also for cell labeling, imaging, and diagnosis.
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影响因子：
3.6
作者：
Freise AC;Wu AM
通讯作者：
Wu AM
DOI：
10.1016/j.ddtec.2018.09.004
发表时间：
2018-12-01
期刊：
Drug discovery today. Technologies
影响因子：
--
作者：
Forte, Nafsika;Chudasama, Vijay;Baker, James R
通讯作者：
Baker, James R
影响因子：
15
作者：
Casi, Giulio;Huguenin-Dezot, Nicolas;Neri, Dario
通讯作者：
Neri, Dario
影响因子：
2.9
作者：
Chevreux, Guillaume;Tilly, Nolwenn;Bihoreau, Nicolas
通讯作者：
Bihoreau, Nicolas
影响因子：
4.7
作者：
Badescu, George;Bryant, Penny;Godwin, Antony
通讯作者：
Godwin, Antony