Bujak Emil, Matasci Mattia, Neri Dario, Wulhfard Sarah
Philochem AG, Zurich, Switzerland.
Methods Mol Biol. 2014;1131:315-34. doi: 10.1007/978-1-62703-992-5_20.
The scFv-Fc format allows for rapid characterization of candidate scFvs isolated from phage display libraries before conversion into a full-length IgG. This format offers several advantages over the phage display-derived scFv, including bivalent binding, longer half-life, and Fc-mediated effector functions. Here, a detailed method is presented, which describes the cloning, expression, and purification of an scFv-Fc fragment, starting from scFv fragments obtained from a phage display library. This method facilitates the rapid screening of candidate antibodies, prior to a more time-consuming conversion into a full IgG format. Alternatively, the scFv-Fc format may be used in the clinic for therapeutic applications.
单链抗体-免疫球蛋白融合蛋白（scFv-Fc）形式能够在将从噬菌体展示文库中分离出的候选单链抗体（scFv）转化为全长免疫球蛋白G（IgG）之前，对其进行快速表征。这种形式相对于噬菌体展示来源的单链抗体具有多个优势，包括二价结合、更长的半衰期以及Fc介导的效应功能。本文介绍了一种详细方法，该方法描述了从噬菌体展示文库获得的单链抗体片段开始，单链抗体-免疫球蛋白融合蛋白片段的克隆、表达和纯化。此方法有助于在将其转化为更耗时的全长IgG形式之前，对候选抗体进行快速筛选。另外，单链抗体-免疫球蛋白融合蛋白形式可用于临床治疗应用。
