Kotlan Beatrix, Glassy Mark C
Integrated Medical Sciences Association Foundation, San Diego, CA, USA.
Methods Mol Biol. 2009;562:1-15. doi: 10.1007/978-1-60327-302-2_1.
Antibody-based immunologic reagents are useful for identifying, isolating, or eliminating cells with particular characteristics related to different diseases. Phage display is a highly valuable technique for antibody selection related to this purpose. In brief, a diverse group of antibody genes prepared from a patient or generated in vitro are inserted into a phagemid vector or the phage genome so that when the protein is expressed, it becomes anchored on the surface of the phage by fusion to a coat protein. A diverse library of recombinant antibodies is generated in this way and can then be exposed or panned on the antigen of interest, typically, this being a molecule associated with a particular pathological condition. Phage that carry proteins or peptides bind preferentially to the target and can thus be isolated from the library. The viruses that are recovered in this way also carry the gene for the binding moiety facilitating its over-expression or manipulation. Recent reviews highlight key milestones in the development of antibody libraries and their screening by phage display, and the impact of these technologies on drug discovery seems assured.
基于抗体的免疫试剂可用于识别、分离或消除具有与不同疾病相关的特定特征的细胞。噬菌体展示是用于与此目的相关的抗体筛选的一项极有价值的技术。简而言之,从患者体内制备或体外产生的一组多样化的抗体基因被插入到噬菌粒载体或噬菌体基因组中,这样当蛋白质表达时,它通过与外壳蛋白融合而锚定在噬菌体表面。通过这种方式产生了一个多样化的重组抗体文库,然后可以使其与感兴趣的抗原接触或淘选,通常该抗原是与特定病理状况相关的分子。携带蛋白质或肽的噬菌体优先与靶标结合,因此可以从文库中分离出来。以这种方式回收的病毒还携带结合部分的基因,便于其过量表达或操作。最近的综述突出了抗体文库开发及其通过噬菌体展示进行筛选的关键里程碑,并且这些技术对药物发现的影响似乎是确定无疑的。