Abstract
The label free real time surface plasmon resonance based immuno-sensing system was first time demonstrated in the authors’ lab using a mercaptoundecanoic acid modified gold sensor chip. The kinetics of the antigen–antibody interaction was determined for optimization of resonance signal intensity for the detection of antigen(s) of fungal teliospores-causative entities of Karnal bunt disease of wheat incited by Tilletia indica. The approach involves the use of a rabbit polyclonal (anti-teliospore) antibody and mono-specific antibody generated against intact teliospores and purified diagnostic antigen of 28 kDa protein. The sensitivity of each of these two immunosensors constructed with the different concentrations of teliosporic antigen was determined by sensogram analysis which showed detection sensitivity as low as 625 and 156 pg which is equivalent to 2.5 and 1.0 teliospores over the sensor surface by getting an angle of dip indicating greater affinity of raised anti-teliospore antibodies and monospecific antibody respectively. By using kinetic evaluation software, the equilibrium constant (KD) and maximum binding capacity of analyte (Bmax) values were calculated and found to be 3.45 and 331.8 nM respectively using polyclonal and 1.57 and 289.57 nM respectively using monospecific antibody.
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Schedule a callMore From: Proceedings of the National Academy of Sciences, India Section B: Biological Sciences
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