Autodisplay of streptavidin

被引：27

作者：

Park, Min ^{[1
]}

Jose, Joachim ^{[2
]}

Thoemmes, Sarah ^{[2
]}

Kim, Jo-Il ^{[1
]}

Kang, Min-Jung ^{[3
]}

Pyun, Jae-Chul ^{[1
]}

机构：

[1] Yonsei Univ, Coll Engn, Sch Mat & Sci, Seoul 120749, South Korea

[2] Univ Dusseldorf, Inst Pharmaceut Chem, Dusseldorf, Germany

[3] Korea Inst Sci & Technol, Seoul, South Korea

来源：

ENZYME AND MICROBIAL TECHNOLOGY | 2011年 / 48卷 / 4-5期

关键词：

Autodisplay; Streptavidin; E; coli; Immunoassay; BACTERIAL SURFACE DISPLAY; ESCHERICHIA-COLI; PROTEIN-A; ANTIBODY IMMOBILIZATION; OUTER-MEMBRANE; AUTOTRANSPORTER PATHWAY; RECOMBINANT PROTEINS; Z-DOMAIN; AIDA-I; AVIDIN;

D O I：

10.1016/j.enzmictec.2010.12.006

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Streptavidin was expressed on the outer membrane of E. coli as a recombinant fusion protein with an autotransporter domain called AIDA-I (adhesin involved in diffuse adherence) using autodisplay technology. The autodisplay of streptavidin was confirmed by SOS-PAGE of the outer membrane proteins, and the number of autodisplayed streptavidin molecules on a single E. coli cell was evaluated with densitometric analysis. The biotin-binding activity of the autodisplayed streptavidin was estimated after treatment with fluorescently labeled biotin by fluorescence microscopy and flow cytometry. The biotin-binding activity of the E. coli with autodisplayed streptavidin was compared with the activity of streptavidin immobilized on magnetic beads. Finally, the outer membrane presenting autodisplayed streptavidin was isolated and layered on a 96-well microplate for an immunoassay. (C) 2010 Elsevier Inc. All rights reserved.

引用

页码：307 / 311

页数：5

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