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    • 2. 发明申请
    • HUMANIZATION AND AFFINITY-OPTIMIZATION OF ANTIBODIES
    • 抗体的人类和亲和力优化
    • WO2010056893A1
    • 2010-05-20
    • PCT/US2009/064247
    • 2009-11-12
    • IMCLONE LLCHAIDAR, Jaafar, Nassar SleimanYUAN, Qing-anZHU, Zhenping
    • HAIDAR, Jaafar, Nassar SleimanYUAN, Qing-anZHU, Zhenping
    • C07K16/46C12P21/08G01N33/48
    • C07K16/464C07K16/005C07K16/2863C07K2317/24C07K2317/55
    • In an efficient selection of higher affinity binders for an antibody humanization strategy, original CDRs are grafted, but no homology modeling is required to determine the key residues in a framework region. One human variable region heavy chain or "VH' and one variable region light chain or 'VL" germline segments show characteristics of high expression in E. coli as a universal template. Key residues in the framework scaffold were determined by screening of available antibody crystal structure data as well as published documents that described the refinements of a specific mouse humanization project. Diversified residues that represent either human antibody selections or mouse selections were integrated into those key framework positions that were assumed to be important in either supporting CDRs or affecting the overall domain structure. A phage display Fab library was used to screen functional reshaped antibodies, which facilitates the isolation of well-maintained clones in a standard panning.
    • 在用于抗体人源化策略的高亲和力结合物的有效选择中,原始CDR被移植,但是不需要同源建模来确定框架区域中的关键残基。 一个人可变区重链或“VH”和一个可变区轻链或“VL”种系显示了在大肠杆菌中作为通用模板的高表达特征。 通过筛选可获得的抗体晶体结构数据以及描述特定小鼠人源化项目的改进的公开文献来确定框架支架中的关键残基。 代表人抗体选择或小鼠选择的多样化残基被整合到那些被认为在支持CDR或影响整个结构域结构中重要的关键框架位置。 使用噬菌体展示Fab文库筛选功能重整抗体,这有助于在标准淘选中分离良好维持的克隆。