Antibody phage display has been pivotal in the quest to generate human monoclonal antibodies for biomedical and research applications. Target antigen preparation is a main bottleneck associated with the panning process. This includes production complexity, downstream purification, quality and yield. In many instances, purified antigens are preferred for panning but this may not be possible for certain difficult target antigens. Here, we describe an improved procedure of affinity selection against crude or non-purified antigen by saturation of non-binders with blocking agents to promote positive binder enrichment termed as Yin-Yang panning. A naïve human scFv library with kappa light chain repertoire with a library size of 10⁹ was developed. The improved Yin-Yang biopanning process was able to enrich monoclonal antibodies specific to the MERS-CoV nucleoprotein. Three unique monoclonal antibodies were isolated in the process. The Yin-Yang biopanning method highlights the possibility of utilizing crude antigens for the isolation of monoclonal antibodies by phage display.

抗体噬菌体展示对于生成用于生物医学和研究应用的人单克隆抗体至关重要。靶抗原制备是与淘选过程相关的主要瓶颈。这包括生产复杂性、下游纯化、质量和产量。在许多情况下，纯化的抗原优选用于淘选，但这对于某些困难的靶抗原来说可能是不可能的。在这里，我们描述了一种针对粗制或未纯化抗原的亲和力选择的改进程序，通过用封闭剂饱和非结合剂来促进阳性结合剂富集，称为阴阳淘选。开发了一个具有 kappa 轻链库的天然人类 scFv 文库，文库大小为 10 ^{9 。}改进的阴阳生物淘选过程能够富集针对 MERS-CoV 核蛋白的单克隆抗体。在此过程中分离出三种独特的单克隆抗体。阴阳生物淘选方法强调了利用粗抗原通过噬菌体展示分离单克隆抗体的可能性。