I'm developing multiplexed bead based indirect immunoassay for four target cytokines and developing sandwich immunoassay for each. I have measured the performance of each individually and I am evaluating their performance in multiplexed form. While I am clear about the experimental design to evaluate this at different levels( capture Ab, cytokine,and detection Ab) However, I'm looking for the appropriate way to calculate the cross reactivity values.
Georg Pongratz
Intuitive answer:
Antibodies: A,B,C,D
Antigens:a,b,c,d,
E.g. crossreactivity for a: use following two setups: 1. ABCD+a; 2. BCD+a and calculate crossreactivity (%): 100* (ABCD+a - BCD+a) / ABCD+a
repeat for b,c,d.
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