通过 Sortase A 和 Butelase 1 的组合进行 IgG1 的一锅双标记和 C-C 融合蛋白的制备。
One-Pot Dual Labeling of IgG 1 and Preparation of C-to-C Fusion Proteins Through a Combination of Sortase A and Butelase 1.
机构信息
Program in Cellular and Molecular Medicine , Boston Children's Hospital, Harvard Medical School , Boston , Massachussets 02115 , United States.
Department of Microbiology , Massachusetts Institute of Technology , Cambridge , Massachussets 02139 , United States.
出版信息
Bioconjug Chem. 2018 Oct 17;29(10):3245-3249. doi: 10.1021/acs.bioconjchem.8b00563. Epub 2018 Sep 21.
Abstract
Site-specific chemical modification of proteins can assist in the study of their function. Furthermore, these methods are essential to develop biologicals for diagnostic and therapeutic use. Standard protein engineering protocols and recombinant expression enable the production of proteins with short peptide tags recognized by enzymes capable of site-specific modification. We report here the application of two enzymes of orthogonal specificity, sortase A and butelase 1, to prepare non-natural C-to-C fusion proteins. Using these enzymes, we further demonstrate site-selective installation of different chemical moieties at two sites in a full-size antibody molecule.
摘要
蛋白质的定点化学修饰可以辅助其功能研究。此外，这些方法对于开发用于诊断和治疗的生物制剂也至关重要。标准的蛋白质工程方案和重组表达使带有短肽标签的蛋白质得以生产，这些标签可被能够进行定点修饰的酶识别。我们在此报告了两种正交特异性酶（即 sortase A 和 butelase 1）在制备非天然 C-C 融合蛋白中的应用。利用这些酶，我们进一步在一个完整抗体分子的两个位点展示了化学基团的位点选择性连接。
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