Benhar I
Department of Molecular Microbiology and Biotechnology, The George S. Wise Faculty of Life Sciences, Green Building, Room 202, Tel-Aviv University, Ramat Aviv 69978, Israel.
Biotechnol Adv. 2001 Feb 1;19(1):1-33. doi: 10.1016/s0734-9750(00)00054-9.
In recent years, the use of surface-display vectors for displaying polypeptides on the surface of bacteriophage and bacteria, combined with in vitro selection technologies, has transformed the way in which we generate and manipulate ligands, such as enzymes, antibodies and peptides. Phage display is based on expressing recombinant proteins or peptides fused to a phage coat protein. Bacterial display is based on expressing recombinant proteins fused to sorting signals that direct their incorporation on the cell surface. In both systems, the genetic information encoding for the displayed molecule is physically linked to its product via the displaying particle. Using these two complementary technologies, we are now able to design repertoires of ligands from scratch and use the power of affinity selection to select those ligands having the desired (biological) properties from a large excess of irrelevant ones. With phage display, tailor-made proteins (fused peptides, antibodies, enzymes, DNA-binding proteins) may be synthesized and selected to acquire the desired catalytic properties or affinity of binding and specificity for in vitro and in vivo diagnosis, for immunotherapy of human disease or for biocatalysis. Bacterial surface display has found a range of applications in the expression of various antigenic determinants, heterologous enzymes, single-chain antibodies, and combinatorial peptide libraries. This review explains the basis of phage and bacterial surface display and discusses the contributions made by these two leading technologies to biotechnological applications. This review focuses mainly on three areas where phage and cell display have had the greatest impact, namely, antibody engineering, enzyme technology and vaccine development.
近年来,利用表面展示载体在噬菌体和细菌表面展示多肽,并结合体外筛选技术,已经改变了我们生成和操控配体(如酶、抗体和肽)的方式。噬菌体展示基于表达与噬菌体外壳蛋白融合的重组蛋白或肽。细菌展示基于表达与分选信号融合的重组蛋白,这些分选信号指导它们整合到细胞表面。在这两种系统中,编码展示分子的遗传信息通过展示颗粒与其产物物理相连。利用这两种互补技术,我们现在能够从头设计配体文库,并利用亲和力选择的力量从大量无关配体中选择具有所需(生物学)特性的配体。通过噬菌体展示,可以合成并选择定制的蛋白质(融合肽、抗体、酶、DNA结合蛋白),以获得所需的催化特性或结合亲和力以及体外和体内诊断、人类疾病免疫治疗或生物催化的特异性。细菌表面展示已在各种抗原决定簇、异源酶、单链抗体和组合肽文库的表达中得到一系列应用。本文综述解释了噬菌体和细菌表面展示的基础,并讨论了这两种领先技术对生物技术应用的贡献。本文综述主要关注噬菌体和细胞展示产生最大影响的三个领域,即抗体工程、酶技术和疫苗开发。