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Transposition-mediated identification of specific binding or functional proteins

机译:转座介导的特异性结合蛋白或功能蛋白的鉴定

摘要

The method disclosed herein describes a novel technology offering unparalleled efficiency, flexibility, utility and speed for the discovery and optimization of polypeptides having desired binding specificity and/or functionality, including antigen-binding molecules such as antibodies and fragments thereof, for desired functional and/or binding phenotypes. The novel method is based on transposable constructs and diverse DNA libraries cloned into transposable vectors and their transfection into host cells by concomitant transient expression of a functional transposase enzyme. This ensures an efficient, stable introduction of the transposon-based expression vectors into vertebrate host cells in one step, which can then be screened for a desired functional or binding phenotype of the expressed proteins, after which the relevant coding sequences for the expressed proteins, including antibodies and fragments thereof, can be identified by standard cloning and DNA sequencing techniques.
机译:本文公开的方法描述了一种新颖的技术,其为发现和优化具有期望的结合特异性和/或功能的多肽(包括诸如抗体及其片段的抗原结合分子)提供了无与伦比的效率,灵活性,实用性和速度,以实现期望的功能和/或或结合表型。该新方法基于转座构建体和克隆到转座载体中的各种DNA库,并通过功能性转座酶的瞬时表达将它们转染到宿主细胞中。这样可确保将基于转座子的表达载体高效,稳定地导入脊椎动物宿主细胞,然后可以对其进行筛选,以筛选出所需表达蛋白的功能或结合表型,然后对表达蛋白的相关编码序列进行筛选,可以通过标准克隆和DNA测序技术鉴定包括抗体及其片段在内的抗体。

著录项

  • 公开/公告号US10174309B2

    专利类型

  • 公开/公告日2019-01-08

    原文格式PDF

  • 申请/专利权人 NBE-THERAPEUTICS LLC;

    申请/专利号US201314414981

  • 发明设计人 ULF GRAWUNDER;

    申请日2013-07-18

  • 分类号C12N1/06;C12N15/10;C12N5/10;G01N33/68;C07K16/28;C12N9/12;

  • 国家 US

  • 入库时间 2022-08-21 12:04:34

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