Forced Degradation Testing as Complementary Tool for Biosimilarity Assessment

• Published in: Bioengineering (Basel) Vol. 6; no. 3; p. 62
• Main Authors: Dyck, Yan Felix Karl, Rehm, Daniel, Joseph, Jan Felix, Winkler, Karsten, Sandig, Volker, Jabs, Wolfgang, Parr, Maria Kristina
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 21.07.2019; MDPI
• Subjects: Antibodies; Antigens; Bevacizumab; Bioengineering; Biological products; Biopharmaceuticals; biosimilar; forced stability testing; Humidity; Hydrogen peroxide; Infliximab; Investigations; Liquid chromatography; liquid chromatography-mass spectrometry (LC-MS); Mass spectrometry; Mass spectroscopy; Methionine; middle-up approach; Monoclonal antibodies; Oxidation; Oxidative stress; Peptides; Proteins; QTOF-MS; Quality control; Quality management; structure reactivity relationship; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Vascular endothelial growth factor; Germany; QTOF-MS; forced stability testing; middle-up approach; infliximab; liquid chromatography-mass spectrometry (LC-MS); structure reactivity relationship; bevacizumab; biopharmaceuticals; biosimilar
• ISSN: 2306-5354; 2306-5354
• DOI: 10.3390/bioengineering6030062

Oxidation of monoclonal antibodies (mAbs) can impact their efficacy and may therefore represent critical quality attributes (CQA) that require evaluation. To complement classical CQA, bevacizumab and infliximab were subjected to oxidative stress by H O for 24, 48, or 72 h to probe their oxidation susceptibility. For investigation, a middle-up approach was used utilizing liquid chromatography hyphenated with mass spectrometry (LC-QTOF-MS). In both mAbs, the Fc/2 subunit was completely oxidized. Additional oxidations were found in the light chain (LC) and in the Fd' subunit of infliximab, but not in bevacizumab. By direct comparison of methionine positions, the oxidized residues in infliximab were assigned to M55 in LC and M18 in Fd'. The forced oxidation approach was further exploited for comparison of respective biosimilar products. Both for bevacizumab and infliximab, comparison of posttranslational modification profiles demonstrated high similarity of the unstressed reference product (RP) and the biosimilar (BS). However, for bevacizumab, comparison after forced oxidation revealed a higher susceptibility of the BS compared to the RP. It may thus be considered a useful tool for biopharmaceutical engineering, biosimilarity assessment, as well as for quality control of protein drugs.