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Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element
Özlem Ertekin1,2, Selma Öztürk3, Zafer Ziya Öztürk4
1TÜBİTAK, The Scientific and Technological Research Council of Turkey, Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, 41470 Kocaeli, Turkey. ozlem.ertekin@tubitak.gov.tr.
Sensors (Basel, Switzerland)
|August 17, 2016
Summary
This study developed a sensitive Quartz Crystal Microbalance (QCM) immunobiosensor for aflatoxin (AF) detection. The IgA antibody demonstrated a higher detection range than IgG antibodies in this novel AF biosensor.
Area of Science:
- Biotechnology
- Biosensor Development
- Immunotechnology
Background:
- Aflatoxins (AF) are toxic secondary metabolites posing significant health risks.
- Development of sensitive and reliable biosensors for AF detection is crucial for food safety.
- Existing immunobiosensors often utilize IgG antibodies, with limited exploration of IgA's potential.
Purpose of the Study:
- To develop a highly sensitive Quartz Crystal Microbalance (QCM) immunobiosensor for aflatoxin detection.
- To evaluate the performance of IgA isotype monoclonal antibodies compared to IgG antibodies in an inhibitory immunoassay format.
- To optimize sensor surface preparation for enhanced stability and reduced non-specific binding.
Main Methods:
- Development of a QCM immunobiosensor using covalent immobilization of AFB1 onto a gold-coated Quartz Crystal via self-assembled monolayer (SAM) and EDC/NHS chemistry.
- Comparative analysis of IgA and IgG monoclonal antibodies with similar aflatoxin affinity.
- Optimization of sensor surface preparation, including minimizing EDC/NHS activation time and chemical blocking to eliminate protein blocking.
- Testing sensor chip stability in harsh solutions for regeneration of antibody-antigen interaction.
Main Results:
- The IgA isotype antibody demonstrated a higher detection range compared to the IgG antibody in the developed QCM immunosensor for AFB1.
- The higher molecular weight of IgA antibodies proved advantageous for label-free immunobiosensor measurements.
- The protein-free sensor chip exhibited excellent stability in harsh solutions, allowing for regeneration.
Conclusions:
- IgA antibodies offer superior performance over IgG antibodies for aflatoxin detection in QCM immunobiosensors.
- The developed IgA-based QCM immunobiosensor provides a sensitive and robust platform for aflatoxin monitoring.
- The protein-free sensor chip design enhances durability and reusability, crucial for practical applications.
