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SCREENING AND ENGINEERING METHOD OF SUPER-STABLE IMMUNOGLOBULIN VARIABLE DOMAINS AND THEIR USES

机译:超稳定免疫球蛋白可变区的筛选与工程方法及其应用

摘要

There are provided a method named Tat-associated protein engineering (TAPE), of screening a target protein having higher solubility and excellent thermostability, in particular, an immunoglobulin variable domain (VH or VL) derived from human germ cells, by preparing a gene construct where the target protein and an antibioticresistant protein are linked to a Tat signal sequence, and then expressing this within E . coli, and human or engineered VH and VL domain antibodies and human or engineered VH and VL domain antibody scaffolds having solubility and excellent thermostability, which are screened by the TAPE method. There are also provided a library including random CDR sequences in the human or engineered VH or VL domain antibody scaffold screened by the TAPE method, and a preparing method thereof. There are also provided a VH or VL domain antibody having binding ability to the target protein screened by using the library, and a pharmaceutical composition including the domain antibody.
机译:提供了一种称为Tat相关蛋白工程(TAPE)的方法,其通过制备基因构建体来筛选具有更高溶解度和优异的热稳定性的靶蛋白,特别是来源于人生殖细胞的免疫球蛋白可变域(VH或VL)。其中靶蛋白和抗生素抗性蛋白与Tat信号序列连接,然后在E.中表达。通过TAPE方法筛选的具有溶解性和优异的热稳定性的大肠杆菌,人或工程VH和VL结构域抗体以及人或工程VH和VL结构域抗体支架。还提供了包含通过TAPE方法筛选的人或工程改造的VH或VL结构域抗体支架中的随机CDR序列的文库及其制备方法。还提供了具有通过使用文库筛选的与靶蛋白的结合能力的VH或VL结构域抗体,以及包含该结构域抗体的药物组合物。

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