Kinetic Binding Analysis of Biomolecular Interactions Using Surface Plasmon Resonance System

K. Narsaiah; S.N. Jha; Pranita Jaiswal; Ashish Kumar Singh; Poonam Preet Kaur; Rajiv Sharma; Ramesh Kumar

doi:10.52151/jae2016532.1602

Authors

K. Narsaiah ICAR-Central Institute of Post-harvest Engineering and Technology, Ludhiana-141004, Punjab, India Author
S.N. Jha Agrie. Engineering Division, ICAR, KAB II, New Delhi-110012 Author
Pranita Jaiswal Agriculture Structures and Environmental Control Division, ICAR-Central Institute of Post-harvest Engineering and Technology, Ludhiana-141004, Punjab, India Author
Ashish Kumar Singh Agriculture Structures and Environmental Control Division, ICAR-Central Institute of Post-harvest Engineering and Technology, Ludhiana-141004, Punjab, India Author
Poonam Preet Kaur Agriculture Structures and Environmental Control Division, ICAR-Central Institute of Post-harvest Engineering and Technology, Ludhiana-141004, Punjab, India Author
Rajiv Sharma Agriculture Structures and Environmental Control Division, ICAR-Central Institute of Post-harvest Engineering and Technology, Ludhiana-141004, Punjab, India Author
Ramesh Kumar Horticulture Crop Processing Division, ICAR-Central Institute of Post-harvest Engineering and Technology, Abohar-152116, Punjab, India. Author

DOI:

https://doi.org/10.52151/jae2016532.1602

Keywords:

Antigen, antibody, small molecule, biomolecular interaction, binding kinetics, enzyme inhibition

Abstract

Surface plasmon resonance (SPR) technology is a versatile and sensitive tool widely used for kinetic studies of interactions between unlabelled biomolecules in real time. The interactions of 4-carboxy benzene sulfonamide (4-CBS) inhibitor, with the enzyme carbonic anhydrase II (CA II) and Interleukin 2 with its antibody (IL 2 antibody) were determined at different concentrations by injecting at a flow rate of 100 μl.min-1 for 60s (100 μl) for a 1 min association phase, which was followed by a 2 min dissociation phase. Langmuir model yielded the rate constants ka and kd , which were of the order of 1.58E+3 M-1. s-1 and 0.04 s-1, respectively, and equilibrium dissociation constant was 2.45E-7 M. Automatic artifact removal, using software options and double referencing (by injection of running buffer to nullify non-specific interactions), further improved the quality of response. Langmuir model yielded the rate constants ka and kd , which were of the order of 2.79E+5 M-1.s-1 and 1.43E-4 s-1, respectively, and equilibrium dissociation constant was 5.17E-7 M. The results illustrate the potential of such kinetic analysis in disease management, drug discovery and for detection of pathogens, toxins, pesticides and antibiotic residues based on either enzyme inhibition or antigen-antibody binding

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