The use of micro Pillar Array Columns (μPACᵀᴹ) for monoclonal antibody (mAb) peptide mapping is presented. Remicade® (infliximab) 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, minute differences such as point mutations could be detected in the protein sequence. According to US and European regulatory authorities identical primary sequence is primordial to similarity. Using μPACᵀᴹ as a tool in biosimilar characterization could therefore be of great value as demonstrated by these results.