Monoclonal antibody peptide mapping – Herceptine® originator and candidate biosimilar

The use of micro Pillar Array Columns (μPACTM) for monoclonal antibody (mAb) peptide mapping is presented. Herceptine® (trastuzumab) originator and candidate biosimilar 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).

Using this high-resolution and high sensitivity separation process, post-translational modifications such as glycosylation, asparagine deamidation, aspartate isomerization, oxidation, N-terminal cyclization (pyroglutamate) and C-terminal lysine truncation, amongst others were revealed. This makes μPACTM technology attractive to compare different mAb production batches and to compare mAb originator products to biosimilars.