Parameters affecting the display of antibodies on phage

被引：39

作者：

Kirsch, M ^{[1
]}

Zaman, M ^{[1
]}

Meier, D ^{[1
]}

Dübel, S ^{[1
]}

Hust, M ^{[1
]}

机构：

[1] Tech Univ Carolo Wilhelmina Braunschweig, Abt Biotechnol, Inst Biochem & Biotechnol, D-38106 Braunschweig, Germany

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 2005年 / 301卷 / 1-2期

关键词：

Fab; phage display; recombinant antibody; scFv; vector design;

D O I：

10.1016/j.jim.2005.04.017

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Despite the fact that a multitude of antibody phage display libraries has been built, systematic comparisons of critical design parameters are rare. Here we analysed the impact of various factors on the performance of the phage display system. First, we compared several vector designs for the display of Fab fragments of antibodies. Bicistronic as well as monocistronic expression of the antibody/pIII operon and vectors using fd-pIII as well as LC-pIII fusions were tested. Further, we evaluated the influence of glucose on the promoter induction. We compared monovalent versus oligovalent display of the antibody fragments and we used antibody fragments with different folding efficiency to assess the influence of the individual antibody sequences on the performance of the system. Finally, both phage display efficiency and yield of soluble Fab fragments were analysed. The significant differences found for phage yield, display of Fabs on the phage and expression of soluble Fabs suggest to use a bicistronic vector with an fd-fragment-pIII fusion for the construction of future Fab phage display libraries. (c) 2005 Elsevier B.V. All rights reserved.

引用

页码：173 / 185

页数：13

共 31 条

[1] An improved helper phage system for efficient isolation of specific antibody molecules in phage display [J].