Ma H, O'Kennedy R
Biomedical Diagnostics Institute, Dublin City University, Dublin 9, Ireland.
Biomedical Diagnostics Institute, Dublin City University, Dublin 9, Ireland.
Methods. 2017 Mar 1;116:23-33. doi: 10.1016/j.ymeth.2016.11.008. Epub 2016 Nov 18.
Recombinant antibodies are now very important in both therapeutics and diagnostics and offer significant advantages over conventional antibodies. The generation of a single-chain variable antibody fragment (scFv) (a common and important recombinant antibody format) is used to demonstrate the construction of a recombinant antibody library. An immunotube-based two-day panning approach, using Escherichia coli as an expression system, is utilised for antibody screening. The methods used for antibody selection and purification using immobilised metal affinity chromatography (IMAC) are described.
重组抗体如今在治疗学和诊断学领域都非常重要,并且相较于传统抗体具有显著优势。单链可变抗体片段(scFv,一种常见且重要的重组抗体形式)的生成用于展示重组抗体文库的构建。采用基于免疫管的为期两天的淘选方法,以大肠杆菌作为表达系统进行抗体筛选。还描述了使用固定化金属亲和色谱(IMAC)进行抗体选择和纯化的方法。