Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element
This study introduces the use of an IgA isotype aflatoxin (AF) specific monoclonal antibody for the development of a highly sensitive Quartz Crystal Microbalance (QCM) immunobiosensor for the detection of AF in inhibitory immunoassay format. The higher molecular weight of IgA antibodies proved an ad...
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doaj-4d4269215c88422c9a60a6a29293cd692020-11-24T21:48:17ZengMDPI AGSensors1424-82202016-08-01168127410.3390/s16081274s16081274Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition ElementÖzlem Ertekin0Selma Öztürk1Zafer Ziya Öztürk2TÜBİTAK, The Scientific and Technological Research Council of Turkey, Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, 41470 Kocaeli, TurkeyTÜBİTAK, The Scientific and Technological Research Council of Turkey, Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, 41470 Kocaeli, TurkeyDepartment of Physics, Gebze Technical University, 41400 Kocaeli, TurkeyThis study introduces the use of an IgA isotype aflatoxin (AF) specific monoclonal antibody for the development of a highly sensitive Quartz Crystal Microbalance (QCM) immunobiosensor for the detection of AF in inhibitory immunoassay format. The higher molecular weight of IgA antibodies proved an advantage over commonly used IgG antibodies in label free immunobiosensor measurements. IgA and IgG antibodies with similar affinity for AF were used in the comparative studies. Sensor surface was prepared by covalent immobilization of AFB1, using self assembled monolayer (SAM) formed on gold coated Quartz Crystal, with 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxy succinimide (EDC/NHS) method using a diamine linker. Nonspecific binding to the surface was decreased by minimizing the duration of EDC/NHS activation. Sensor surface was chemically blocked after AF immobilization without any need for protein blocking. This protein free sensor chip endured harsh solutions with strong ionic detergent at high pH, which is required for the regeneration of the high affinity antibody-antigen interaction. According to the obtained results, the detection range with IgA antibodies was higher than IgG antibodies in QCM immunosensor developed for AFB1.http://www.mdpi.com/1424-8220/16/8/1274QCM immunosensorIgA monoclonal antibodyaflatoxin immobilizationchemical blocking |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Özlem Ertekin Selma Öztürk Zafer Ziya Öztürk |
spellingShingle |
Özlem Ertekin Selma Öztürk Zafer Ziya Öztürk Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element Sensors QCM immunosensor IgA monoclonal antibody aflatoxin immobilization chemical blocking |
author_facet |
Özlem Ertekin Selma Öztürk Zafer Ziya Öztürk |
author_sort |
Özlem Ertekin |
title |
Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element |
title_short |
Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element |
title_full |
Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element |
title_fullStr |
Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element |
title_full_unstemmed |
Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element |
title_sort |
label free qcm immunobiosensor for afb1 detection using monoclonal iga antibody as recognition element |
publisher |
MDPI AG |
series |
Sensors |
issn |
1424-8220 |
publishDate |
2016-08-01 |
description |
This study introduces the use of an IgA isotype aflatoxin (AF) specific monoclonal antibody for the development of a highly sensitive Quartz Crystal Microbalance (QCM) immunobiosensor for the detection of AF in inhibitory immunoassay format. The higher molecular weight of IgA antibodies proved an advantage over commonly used IgG antibodies in label free immunobiosensor measurements. IgA and IgG antibodies with similar affinity for AF were used in the comparative studies. Sensor surface was prepared by covalent immobilization of AFB1, using self assembled monolayer (SAM) formed on gold coated Quartz Crystal, with 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxy succinimide (EDC/NHS) method using a diamine linker. Nonspecific binding to the surface was decreased by minimizing the duration of EDC/NHS activation. Sensor surface was chemically blocked after AF immobilization without any need for protein blocking. This protein free sensor chip endured harsh solutions with strong ionic detergent at high pH, which is required for the regeneration of the high affinity antibody-antigen interaction. According to the obtained results, the detection range with IgA antibodies was higher than IgG antibodies in QCM immunosensor developed for AFB1. |
topic |
QCM immunosensor IgA monoclonal antibody aflatoxin immobilization chemical blocking |
url |
http://www.mdpi.com/1424-8220/16/8/1274 |
work_keys_str_mv |
AT ozlemertekin labelfreeqcmimmunobiosensorforafb1detectionusingmonoclonaligaantibodyasrecognitionelement AT selmaozturk labelfreeqcmimmunobiosensorforafb1detectionusingmonoclonaligaantibodyasrecognitionelement AT zaferziyaozturk labelfreeqcmimmunobiosensorforafb1detectionusingmonoclonaligaantibodyasrecognitionelement |
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