Generation of Thermally Stable Affinity Pairs for Sensitive, Specific Immunoassays.
Generation of Thermally Stable Affinity Pairs for Sensitive, Specific Immunoassays.
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生成热稳定亲和对，用于灵敏、特异性的免疫测定。
DOI：
10.1007/978-1-0716-2285-8_21
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Sikes,HadleyD
中科院分区：
文献类型：
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作者：
Corless,Elliot;Hao,Yining;Jia,Huan;Kongsuphol,Patthara;Tay,DousabelMY;Ng,SayYong;Sikes,HadleyD
许多即时诊断测试依赖于一对单克隆抗体，其结合到感兴趣分子的两个不同表位。该方案描述了基于可替代单克隆抗体的容易产生的小蛋白质支架rcSso7d的这种亲和对的鉴定和产生。这些强结合变体是从大型酵母展示文库中鉴定的。所描述的方法可以显著快于抗体生成和表位分仓，产生在常见细菌蛋白质合成菌株中合成的亲和对，从而能够快速生成新型诊断工具。
Many point-of-care diagnostic tests rely on a pair of monoclonal antibodies that bind to two distinct epitopes of a molecule of interest. This protocol describes the identification and generation of such affinity pairs based on an easily produced small protein scaffold rcSso7d which can substitute monoclonal antibodies. These strong binding variants are identified from a large yeast display library. The approach described can be significantly faster than antibody generation and epitope binning, yielding affinity pairs synthesized in common bacterial protein synthesis strains, enabling the rapid generation of novel diagnostic tools.