I need to immobilize antibodies on a SPRi biochip. I need to optimize the best concentration of antibodies/protein to immobilize on the chip surface. I wants to know what is the best method to determine the concentration of antibodies bound on the chip surface, pre and post concentration determination method. As it is very important key for the quantification purpose through an SPRi experiment.
Thank You
Sincerely Your
You do not need to determine the antibody concentration bound on the chip precisely, if you aim for an antigen quantification. In this case, the calibration is based on a dilution series of antigen. In SPR, the signal is strongly dependent on the distance of a molecule from the surface. Therefore, a quantitative amount of immobilized antibody would not help you much, anyway.
Dear Prof. Thank You so much for your answer. It means I can bind the antibodies with any concentration on chip and just make a dilution series for my sample of target?
Thank You so much
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