Koenig Patrick, Fuh Germaine
Antibody Engineering Department, Genentech Inc., South San Francisco, CA, USA.
Methods Mol Biol. 2014;1131:133-49. doi: 10.1007/978-1-62703-992-5_9.
Phage display is a powerful tool to isolate specific binders from a large and diverse combinatorial library. Here we provide a step-by-step protocol in how to set up a successful phage panning experiment in order to isolate useful antibodies. The protocol includes testing antigens for their suitability in the phage panning procedure and optimizing the panning conditions and alternative screening methods to minimize nonspecific binding. We describe example phage panning experiments starting from the library transformation to the phage clone screening.
噬菌体展示是一种从庞大且多样的组合文库中分离特定结合物的强大工具。在此,我们提供一份关于如何开展成功的噬菌体淘选实验以分离有用抗体的详细步骤方案。该方案包括测试抗原在噬菌体淘选过程中的适用性,优化淘选条件以及采用替代筛选方法以尽量减少非特异性结合。我们描述了从文库转化到噬菌体克隆筛选的噬菌体淘选实验示例。