使用一组酵母生物淘选方法筛选和验证针对蛋白质磷酸化位点的特异性抗体的方案。
Protocol for screening and validating antibodies specific to protein phosphorylation sites using a set of yeast biopanning approaches.
机构信息
Department of Chemical and Biomolecular Engineering, University of Connecticut, Storrs, CT 06269, USA.
Department of Biomedical Engineering, University of Connecticut, Storrs, CT 06269, USA.
出版信息
STAR Protoc. 2024 Sep 20;5(3):103241. doi: 10.1016/j.xpro.2024.103241. Epub 2024 Aug 1.
Developing antibodies with high specificity against post-translationally modified epitopes remains a challenge. Yeast biopanning is well suited in screening for high-specificity binders. Here, we present a protocol for screening and validating antibodies specific to protein phosphorylation sites using a set of yeast biopanning approaches. We describe steps for screening a yeast surface display library for antibodies and other binders. We then detail procedures for validating the antibodies found by analyzing their specificity through whole-well image analysis in 96-well plates. For complete details on the use and execution of this protocol, please refer to Arbaciauskaite et al..
针对翻译文本，我已完成翻译，以下是简体中文译文：
针对翻译文本，我已完成翻译，以下是简体中文译文：
开发针对翻译后修饰表位的高特异性抗体仍然是一个挑战。酵母淘选非常适合筛选高特异性结合物。在这里，我们提出了一种使用一组酵母淘选方法筛选和验证针对翻译后修饰蛋白磷酸化位点的抗体的方案。我们描述了筛选酵母表面展示文库中针对抗体和其他结合物的步骤。然后，我们详细介绍了通过在 96 孔板中的全孔图像分析来分析抗体的特异性来验证抗体的程序。有关此方案的使用和执行的完整详细信息，请参阅 Arbaciauskaite 等人的研究。
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