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Title: A silicon nitride ISFET based immunosensor for Ag85B detection of tuberculosis
Authors: Pawasuth Saengdee
Woraphan Chaisriratanakul
Win Bunjongpru
Witsaroot Sripumkhai
Awirut Srisuwan
Charndet Hruanun
Amporn Poyai
Ponrut Phunpae
Supansa Pata
Wutthinan Jeamsaksiri
Watchara Kasinreak
Chamras Promptmas
Mahidol University
Thailand National Science and Technology Development Agency
Chiang Mai University
Keywords: Biochemistry, Genetics and Molecular Biology;Chemistry;Environmental Science
Issue Date: 21-Oct-2016
Citation: Analyst. Vol.141, No.20 (2016), 5767-5775
Abstract: © 2016 The Royal Society of Chemistry. A silicon nitride Ion Sensitive Field Effect Transistor (ISFET) based immunosensor was developed as a low-cost and label-free electrical detection for the detection of antigen 85 complex B (Ag85B). The sensing membrane of the ISFET was modified with 3-aminopropyltriethoxysilane (APTES) followed by glutaraldehyde (GA), yielding an aldehyde-terminated surface. This group is available for immobilization of a monoclonal antibody against a recombinant Ag85B protein (anti-Ag85B antibody). The optimal concentration for anti-Ag85B antibody immobilization onto the modified ISFET was 100 μg ml-1. This optimal condition provided the maximal binding capability and minimal non-specific background signal. The binding event between the recombinant Ag85B antigen and anti-Ag85B antibody on the ISFET surface is presented by monitoring the gate potential change at a constant drain current. The dose response for the recombinant Ag85B protein showed a linear response between 0.12 and 1 μg ml-1without significant interference from other recombinant proteins. The analytical imprecision (CV%) and accuracy of this Ag85B protein biosensor were 9.73-10.99% and 95.29%, respectively. In addition, an irrelevant antibody and other recombinant proteins were employed as a negative control to demonstrate the non-specific interaction of the antigen and antibody. The success of this immunosensor system for Ag85B protein detection facilitates the construction of a promising device which can shorten the turnaround time for the diagnosis of tuberculosis compared to a standard culture method. Furthermore, this device could also be applied for real-time growth monitoring of Mycobacterium tuberculosis in a mycobacterial culture system.
ISSN: 13645528
Appears in Collections:Scopus 2016-2017

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