Thie Holger, Schirrmann Thomas, Paschke Matthias, Dübel Stefan, Hust Michael
Technische Universität Braunschweig, Institut für Biochemie und Biotechnologie, Abteilung Biotechnologie, Spielmannstr. 7, 38106 Braunschweig, Germany.
N Biotechnol. 2008 Jun;25(1):49-54. doi: 10.1016/j.nbt.2008.01.001. Epub 2008 May 16.
Antibody phage display is a key technology for the generation of recombinant (human) antibodies for research, diagnostics and therapy. Most antibody fragments can only be folded correctly in the oxidizing environment of the periplasm of Escherichia coli. A multitude of leader peptides has been used for secretion of antibody::pIII fusion proteins into the periplasm, but a systematic study of their impact on the performance of antibody phage display systems has not been reported so far. In this work we have analysed the influence of various leader peptides on antibody phage display efficiency and production yields of soluble antibody fragments. Four leader peptides using the Sec pathway (PelB, OmpA, PhoA and pIII) and three using the SRP pathway (DsbA, TorT and TolB) were compared. Both pathways are compatible with antibody phage display and the production of soluble antibody fragments. The applicability of the SRP pathway to antibody phage display and the production of functional scFvs is shown here for the first time.
抗体噬菌体展示是一种用于生成重组（人）抗体以进行研究、诊断和治疗的关键技术。大多数抗体片段只有在大肠杆菌周质的氧化环境中才能正确折叠。众多的前导肽已被用于将抗体::pIII融合蛋白分泌到周质中，但迄今为止尚未见对其对抗体噬菌体展示系统性能影响的系统研究报道。在本研究中，我们分析了各种前导肽对抗体噬菌体展示效率和可溶性抗体片段产量的影响。比较了使用Sec途径的四种前导肽（PelB、OmpA、PhoA和pIII）和使用SRP途径的三种前导肽（DsbA、TorT和TolB）。这两种途径都与抗体噬菌体展示和可溶性抗体片段的产生兼容。本文首次展示了SRP途径在抗体噬菌体展示和功能性单链抗体片段产生中的适用性。
