Hust Michael, Frenzel André, Schirrmann Thomas, Dübel Stefan
Abteilung Biotechnologie, Institut für Biochemie, Biotechnologie und Bioinformatik, Technische Universität Braunschweig, Braunschweig, Germany.
Methods Mol Biol. 2014;1101:305-20. doi: 10.1007/978-1-62703-721-1_14.
Antibodies are indispensable detection reagents for research and diagnostics and represent the biggest class of biological therapeutics on the market. In vitro antibody selection systems offer many advantages over animal-based technologies because the whole selection process is independent of the in vivo immune response. In the last two decades antibody phage display has evolved to the most robust and widely used method and has already yielded thousands of antibodies. The selection of binders by phage display is also referred to as "panning" and based on the specific molecular interaction of antibody phage with an immobilized antigen thus allowing the enrichment and isolation of antigen-specific monoclonal binders from very large antibody gene libraries. Here, we give detailed protocols for the selection of recombinant antibody fragments from antibody gene libraries in microtiter plates.
抗体是研究和诊断中不可或缺的检测试剂,也是市场上最大类别的生物治疗药物。体外抗体筛选系统比基于动物的技术具有许多优势,因为整个筛选过程独立于体内免疫反应。在过去二十年中,抗体噬菌体展示已发展成为最强大且应用最广泛的方法,已经产生了数千种抗体。通过噬菌体展示选择结合物也被称为“淘选”,它基于抗体噬菌体与固定化抗原的特异性分子相互作用,从而能够从非常大的抗体基因库中富集和分离抗原特异性单克隆结合物。在此,我们给出在微孔板中从抗体基因库中筛选重组抗体片段的详细方案。