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

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