A capacitive immunosensor for detection of cholera toxin

被引:60
|
作者
Labib, Mahmoud [1 ,2 ]
Hedstrom, Martin [1 ]
Amin, Magdy [2 ]
Mattiasson, Bo [1 ]
机构
[1] Lund Univ, Dept Biotechnol, S-22100 Lund, Sweden
[2] Cairo Univ, Fac Pharm, Microbial Biotechnol Ctr, Cairo, Egypt
关键词
Cholera toxin; Capacitive immunosensor; Surface plasmon resonance; Enzyme-linked immunosorbent assays; ENDOPLASMIC-RETICULUM; RETROGRADE TRANSPORT; PROTEIN TOXINS; ANTIBODY; SURFACE; BIOSENSOR; GLYCOSPHINGOLIPIDS; IMMUNOASSAY; MONOLAYERS; AFFINITY;
D O I
10.1016/j.aca.2008.12.035
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Contamination of food with biological toxins as well as their potential use as weapons of mass destruction has created an urge for rapid and cost effective analytical techniques capable of detecting trace amounts of these toxins. This paper describes the development of a sensitive method for detection of cholera toxin (CT) using a flow-injection capacitive immunosensor based on self-assembled monolayers. The sensing Surface consists of monoclonal antibodies against the B subunit of CT (anti-CT), immobilized on a gold transducer. Experimental results show that the immunosensor responded linearly to CT concentrations in the range from 1.0 x 10 (13) to 1.0 x 10 (10) M underoptimized conditions.The limit of detection (LOD) was 1.0 x 10 (14) M. Two more analytical methods were employed for detection of CT using the same antibody namely, sandwich ELISA and Surface plasmon resonance (SPR)-based immunosensor. The former had an LOD of 1.2 x 10 (12) M and a working range from 3.7 x 10 (11) to 2.9 x 10 (10) M whereas, the later had an LOD of 1.0 x 10 M and a linearity ranging from 1.0 x 10 (9) to 1.0 x 10 (6) M. These results demonstrate that the developed capacitive inimunosensor system has a higher sensitivity than the other two techniques. The binding affinity of CT to the immobilized anti-CT was determined using the SPR-based immunosensor and an association constant (K-A) of 1.4 x 10(9) M (1) was estimated. (C) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码:255 / 261
页数:7
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