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Kinetics of Binding Interaction between Norfloxacin and Monoclonal Antibody Using Surface Plasmon Resonance

Patamalai Boonserm1, Songchan Puthong2, Sajee Noitang2, Thanaporn Wichai2, Pongsak Khunrae3, Kittinan Komolpis2,4, and Sarintip Sooksai2
1.Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand
2.Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok, Thailand
3.King Mongkut's University of Technology Thonburi, Bangkok, Thailand
4.Food Risk Hub, Research Unit of Chulalongkorn University, Bangkok, Thailand
Abstract—Fluoroquinolones (FQs) are a group of antibiotics which have been used against both Gram-negative and Gram-positive bacteria in domestic animals. Residue of these antibiotics in food products could lead to drug resistance problem in human. As a result, surveillance detection program of these drug residues must be in practice to ensure safety of the consumers. Detections based on immunological method such as Enzyme-Linked Immunosorbent Assay (ELISA) and lateral flow immunoassay or strip test have been widely used as a screening tool in food safety applications. In this method, either an antibody with broad specificity to a group of FQs or several specific antibodies for each antibiotic are required for the assay to cover all FQs of interest. In this study, sensitivity of two monoclonal antibodies, mAb Nor132 and mAb Nor155, against norfloxacin was quantified by an antigen-captured indirect competitive enzyme linked immunosorbent assay. The sensitivity in term of 50% Inhibition Concentration (IC50) of mAb Nor132 and mAb Nor155 was found to be 0.706 μg/ml and 0.072 μg/ml, respectively. Then, the binding interactions of both mAbs and norfloxacin were studied by Surface Plasmon Resonance (SPR). The equilibrium binding constants (KD) of mAb Nor132 and mAb Nor155 were 1.152×10-8 M and 1.996×10-9 M, respectively. The results indicated that sensitivity and affinity of mAb Nor155 were higher than those of mAb Nor132. These suggested that mAb Nor155 was more suitable than mAb Nor132 for using in the development of norfloxacin detection.

Index Terms—norfloxacin, monoclonal antibody, ELISA, surface plasmon resonance, Biacore, antibody detection

Cite:Patamalai Boonserm, Songchan Puthong, Sajee Noitang, Thanaporn Wichai, Pongsak Khunrae, Kittinan Komolpis, and Sarintip Sooksai, "Kinetics of Binding Interaction between Norfloxacin and Monoclonal Antibody Using Surface Plasmon Resonance," International Journal of Pharma Medicine and Biological Sciences, Vol. 9, No. 2, pp. 81-86, April 2020. doi: 10.18178/ijpmbs.9.2.81-86

Copyright © 2020 by the authors. This is an open access article distributed under the Creative Commons Attribution License (CC BY-NC-ND 4.0), which permits use, distribution and reproduction in any medium, provided that the article is properly cited, the use is non-commercial and no modifications or adaptations are made.

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