I am attempting to conjugate an antibody to a maleimide-drug conjugate. All the TCEP reduction protocols I find specify a protein concentration of 1-10 mg/ml in some sort of buffer (sodium borate, or tris or sodium phosphate), ahead of adding a small volume of TCEP. Because I don't have much antibody to play with and I don't want to lose too great a proportion via spin-column and HPLC purification of my ADC, I want to know if I can use a lower initial antibody concentration (eg 0.1mg/ml) and if not, why not? Many thanks!
You would recollect during you laboratory practicals during you graduation days that reduction or conjugation is usually done in MIC of the antibiotic (as resistance or susceptibility (which is the markers for these studies) of the organisms is best seen at that concentrations and any increase in the concentrations might mask these effects) and that is why such low concentrations are used.
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