Hi, I have always done my in-vitro experiments of protein-antibiotic interaction at a pH of 5.5. Now that I want to do some experiments with the SPR, is it still advisable to use this low pH? Thanks!
When you plan to use a dextran based sensor chip the main problem with pH 5.5 is the possible non-specific interaction with the dextran matrix. This is mainly due to free COOH groups that are partially COO- at this pH. You can try other matrices / lower carboxylation surfaces to minimize.
Start with injecting the antibiotic over a new surface at pH 5.5. If there is no binding signal I think your are good to go and try a real binding experiment.
Using immobilization and running buffers at pH 5.5 in Surface Plasmon Resonance (SPR) can have some potential disadvantages.
It is important to consider these potential disadvantages and to test your protein-antibiotic interactions at a range of pH values to determine if pH 5.5 is the optimal condition for your specific system. Additionally, It is also important to check if the immobilization buffer and running buffer that you are using are compatible with the pH you are using.
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