He Mingyue, Taussig Michael J
Technology Research Group, The Babraham Institute, Cambridge, UK.
Methods Mol Biol. 2008;484:193-205. doi: 10.1007/978-1-59745-398-1_13.
Ribosome display is a powerful method for selection of single-chain antibodies in vitro. It operates through the formation of libraries of antibody-ribosome-mRNA complexes that are selected on immobilized antigen, followed by recovery of the genetic information from the mRNA by RT-PCR. Both prokaryotic and eukaryotic versions are used. We describe our eukaryotic system, in which rabbit reticulocyte extracts are used for cell free transcription/translation and cDNA is recovered by in situ RT-PCR performed on the selected complexes.
核糖体展示是一种在体外筛选单链抗体的强大方法。它通过形成抗体-核糖体-mRNA复合物文库来运作，该文库在固定化抗原上进行筛选，随后通过RT-PCR从mRNA中回收遗传信息。原核和真核版本均有使用。我们描述了我们的真核系统，其中兔网织红细胞提取物用于无细胞转录/翻译，并且通过对选定复合物进行原位RT-PCR来回收cDNA。
