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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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抗vstm1-v2细胞因子人源化单链抗体文库的构建及筛选
, PP. 1651-1656
Keywords: vstm1-v2细胞因子,鼠源vh-cdr3,改良的ta克隆,人源化单链抗体
Abstract:
本文将鼠源vh-cdr3库(libraryofmurinevh-cdr3)移植到特定的人源scfv(single-chainantibodyfragment)vh3-vκ1框架构建人源化scfv文库,快速筛选抗vstm1-v2的人源化单链抗体。以抗vstm1-v2鼠源cdna为模板,扩增出鼠源vh-cdr3库,随后将鼠源vh-cdr3库移植到人源scfv(vh3-vκ1)上并通过核糖体展示富集抗vstm1-v2人源化scfv文库,该文库序列通过与原核基因表达调控等成分(n端添加t7启动子、sd序列和pelb序列、c端添加his标签序列和t7terminal序列)拼接,构建表达型ta载体转化大肠杆菌bl21(de3)进行可溶性表达。最终构建了一个库容达1012的人源化单链抗体文库,完成了1000个克隆的初步鉴定,并筛选出ec50达21.35nmol·l-1的抗vstm1-v2人源化scfv。该研究结果,一方面获得了具有潜在应用价值的抗-vstm1-v2先导抗体;另一方面,为快速获得人源化抗体提供了一个新技术途径。
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