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    • 1. 发明申请
    • DIRECTED EVOLUTION OF NOVEL BINDING PROTEINS
    • 新型结合蛋白的指导进化
    • US20070259417A1
    • 2007-11-08
    • US11745199
    • 2007-05-07
    • Robert LadnerSonia GutermanBruce RobertsWilliam MarklandArthur LeyRachel Kent
    • Robert LadnerSonia GutermanBruce RobertsWilliam MarklandArthur LeyRachel Kent
    • C12N1/08
    • C40B40/02A61K8/64A61K38/00A61K2800/57A61Q11/00C07K1/047C07K14/43522C07K14/8114C07K14/8117C12N7/00C12N15/1037C12N2795/18111
    • In order to obtain a novel binding protein against a chosen target, DNA molecules, each encoding a protein comprising one of a family of similar potential binding domains and a structural signal calling for the display of the protein on the outer surface of a chosen bacterial cell, bacterial spore or phage (genetic package) are introduced into a genetic package. The protein is expressed and the potential binding domain is displayed on the outer surface of the package. The cells or viruses bearing the binding domains which recognize the target molecule are isolated and amplified. The successful binding domains are then characterized. One or more of these successful binding domains is used as a model for the design of a new family of potential binding domains, and the process is repeated until a novel binding domain having a desired affinity for the target molecule is obtained. In one embodiment, the first family of potential binding domains is related to bovine pancreatic trypsin inhibitor, the genetic package is M13 phage, and the protein includes the outer surface transport signal of the M13 gene III protein.
    • 为了获得针对所选靶标的新型结合蛋白,每个编码包含相似潜在结合结构域的家族之一的蛋白质的DNA分子和要求在选择的细菌细胞的外表面上显示蛋白质的结构信号 ,细菌孢子或噬菌体(遗传包装)被引入到遗传包装中。 蛋白质被表达,并且潜在的结合结构域显示在包装的外表面上。 分离和扩增具有识别靶分子的结合域的细胞或病毒。 然后表征成功的结合结构域。 将这些成功结合结构域中的一个或多个用作设计新的潜在结合结构域家族的模型,并重复该过程,直到获得对靶分子具有所需亲和力的新结合结构域。 在一个实施方案中,第一潜在结合域家族与牛胰蛋白酶抑制剂有关,遗传包是M13噬菌体,蛋白质包括M13基因III蛋白的外表面转运信号。