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Open AccessArticle

Forced Degradation Testing as Complementary Tool for Biosimilarity Assessment

1
Department of Pharmaceutical & Medicinal Chemistry, Institute of Pharmacy, Freie Universität Berlin, Königin-Luise-Straße 2+4, 14195 Berlin, Germany
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Department of Life Sciences & Technology, Beuth Hochschule für Technik Berlin, Seestraße 64, 13347 Berlin, Germany
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ProBioGen AG, Goethestraße 54, 13086 Berlin, Germany
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Core Facility BioSupraMol, Institute of Pharmacy, Freie Universität Berlin, Königin-Luise-Str. 2+4, 14195 Berlin, Germany
*
Author to whom correspondence should be addressed.
Bioengineering 2019, 6(3), 62; https://doi.org/10.3390/bioengineering6030062
Received: 30 June 2019 / Revised: 15 July 2019 / Accepted: 18 July 2019 / Published: 21 July 2019
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 H2O2 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. View Full-Text
Keywords: middle-up approach; liquid chromatography-mass spectrometry (LC-MS); QTOF-MS; biopharmaceuticals; forced stability testing; structure reactivity relationship; bevacizumab; infliximab; biosimilar middle-up approach; liquid chromatography-mass spectrometry (LC-MS); QTOF-MS; biopharmaceuticals; forced stability testing; structure reactivity relationship; bevacizumab; infliximab; biosimilar
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MDPI and ACS Style

Dyck, Y.F.K.; Rehm, D.; Joseph, J.F.; Winkler, K.; Sandig, V.; Jabs, W.; Parr, M.K. Forced Degradation Testing as Complementary Tool for Biosimilarity Assessment. Bioengineering 2019, 6, 62.

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