As the first step in my reactions for ADC synthesis, I need to buffer exchange the mAb (herceptin) and eliminate histidine (that is present in herceptin) that has cross reaction with my linker. in the next step, I want to become sure that there is not any residual histidine in my mAb. how can I do this with HPLC system? how can I seperate the mAb from histidine? which column and elution buffer should I use? can I do this without derivatization?
Dear Sir. Concerning your issue about how to determine residual histidine in the antibody sample after desalting with HPLC System. Antibodies are members of a family of molecules, the immunoglobulins, that constitute the humoral branch of the immune system and form approximately 20% of the plasma proteins in humans. The solution was separated and dtermined by RP-HPLC with a C8 column. I think the following below links and the attached files may help you in your analysis:
http://www.helixbiopharma.com/wp-content/uploads/2014/02/LDOS47-Bioconjugate-May-2015.pdf
Thanks
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