Hi,
We are working on DAR (Drug to Antibody Ratio) estimation of ADC using the following HIC-UV method. For some of the unconjugated mab we getting a sharp peak but for our desired mab we are getting a broad peak. Do you guess what could be the reason? I have attached a chromatogram for your reference.
Mobile phase A:50mM PO4, pH 7.0, 1.5M (NH4)2SO4, 5%IPA
Mobile phase B:50mM PO4, pH 7.0, 20%IPA
Column: Waters ProteinPak Hi Res HIC 4.6*100mm, 2.5um
Injection vol: 10ul
Do not hesitate to ask if you need more information to understand the case.
Sincerely,
Dhaval