Characterizing antibody-drug conjugates using µPACᵀᴹ-MS

The use of micro pillar array columns (μPACᵀᴹ) for characterizing antibody-drug-conjugates is presented. Kadcyla® (ado-trastuzumab emtansine) and, for comparison, Herceptin® (trastuzumab) tryptic digests were analyzed on a 200 cm μPACᵀᴹ C18 column and peaks eluting were detected by ultraviolet (UV) spectroscopy and high-resolution mass spectrometry (MS).

The resolving power offered by the μPACᵀᴹ column allows an in-depth study of conjugation sites and accommodates the separation of isomeric conjugated peptides. In combination with all-ions MS/MS, conjugates peptides can selectively be recognized facilitating data interpretation.