Hi All,
I am considering raising the concentration of my PBS solution to buffer against a pH drop during a mAb reaction step. Would that increase have any effect of the structure or affinity of my mAb?
Thank you!
EF
Hello Eduard,
Based on my personal experience, dialysing antibodies against 1X PBS and concentrating to 10 fold (final PBS concentration 10X) had no obvious effect on antibody stability/activity. However, in the reaction, the ionic strength of buffer was not that high. I understand you need to determine it empirically. The concentration of phosphate is not high in 1X PBS and I guess increasing the strength by 2-2.5 fold should have little if any effect on antibody-antigen interaction. Alternatively, consider Good's buffers such as HEPES.
It's probably safe to do it a little bit, but if you want to be super-safe, make your own PBS from powdered phosphate & NaCl. Recipes are all over the net. Then, you can raise only the buffer (phosphate) concentration, and drop the NaCl accordingly, to make up for it.
Yes, the commentary is consistent with many other researchers as well as in the Biotechnology industry. In your case it is unlikely you will reach conductivities that are detrimental to human or rodent IgG1 and IgG2 whether it's the phosphate or the NaCl. If you prepare the concentrated buffer from pre-made stocks then be sure that the endotoxin level is checked if you plan for in vivo work particularly for inflammation models.
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