{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T16:15:15Z","timestamp":1784218515923,"version":"3.55.0"},"reference-count":61,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2019,2,27]],"date-time":"2019-02-27T00:00:00Z","timestamp":1551225600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotech &amp; Bioengineering"],"published-print":{"date-parts":[[2019,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Glycosylation, a critical product quality attribute, may affect the efficacy and safety of therapeutic proteins in vivo. Chinese hamster ovary fed\u2010batch cell culture batches yielded consistent glycoprofiles of a Fc\u2010fusion antibody comprizing three different N\u2010glycosylation sites. By adding media supplements at specific concentrations in cell culture and applying enzymatic glycoengineering, a diverse N\u2010glycan variant population was generated, including high mannose, afucosylated, fucosylated, agalactosylated, galactosylated, asialylated, and sialylated forms. Site\u2010specific glycosylation profiles were elucidated by glycopeptide mapping and the effect of the glycosylation variants on the Fc\u03b3RIIIa receptor binding affinity and the biological activity (cell\u2010based and surface plasmon resonance) was assessed. The two fusion body glycosylation sites were characterized by a high degree of sialic acid, more complex N\u2010glycan structures, a higher degree of antennarity, and a site\u2010specific behavior in the presence of a media supplement. On the other hand, the media supplements affected the Fc\u2010site glycosylation heterogeneity similarly to the various studies described in the literature with classical monoclonal antibodies. Enzymatic glycoengineering solely managed to generate high levels of galactosylation at the fusion body sites. Variants with low core fucosylation, and to a lower extent, high mannose glycans exhibited increased Fc\u03b3RIIIa receptor binding affinity. All N\u2010glycan variants exhibited weak effects on the biological activity of the fusion body. Both media supplementation and enzymatic glycoengineering are suitable to generate sufficient diversity to assess the effect of glycostructures on the biological activity.<\/jats:p>","DOI":"10.1002\/bit.26930","type":"journal-article","created":{"date-parts":[[2019,1,19]],"date-time":"2019-01-19T15:02:48Z","timestamp":1547910168000},"page":"1017-1028","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Generation of site\u2010distinct N\u2010glycan variants for in vitro bioactivity testing"],"prefix":"10.1002","volume":"116","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8033-6812","authenticated-orcid":false,"given":"David","family":"Br\u00fchlmann","sequence":"first","affiliation":[{"name":"Merck Biopharma Biotech Process Sciences Fenil\u2010sur\u2010Corsier Switzerland"},{"name":"Department of Biotechnology and Biophysics Julius\u2010Maximilians\u2010Universit\u00e4t W\u00fcrzburg  Biozentrum W\u00fcrzburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Vuillemin","sequence":"additional","affiliation":[{"name":"Merck Biopharma Biotech Process Sciences Fenil\u2010sur\u2010Corsier Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abhijeet","family":"Satwekar","sequence":"additional","affiliation":[{"name":"Merck Serono S.p.A, Analytical Development Biotech Products Guidonia Montecelio Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eugenio","family":"Galano","sequence":"additional","affiliation":[{"name":"Merck Serono S.p.A, Analytical Development Biotech Products Guidonia Montecelio Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angelo","family":"Palmese","sequence":"additional","affiliation":[{"name":"Merck Serono S.p.A, Analytical Development Biotech Products Guidonia Montecelio Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandra","family":"D'Angelo","sequence":"additional","affiliation":[{"name":"Merck Serono S.p.A, Analytical Development Biotech Products Guidonia Montecelio Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zeynep","family":"Manco","sequence":"additional","affiliation":[{"name":"Merck Biopharma Biotech Process Sciences Fenil\u2010sur\u2010Corsier Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan","family":"Souquet","sequence":"additional","affiliation":[{"name":"Merck Biopharma Biotech Process Sciences Fenil\u2010sur\u2010Corsier Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Herv\u00e9","family":"Broly","sequence":"additional","affiliation":[{"name":"Merck Biopharma Biotech Process Sciences Fenil\u2010sur\u2010Corsier Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Markus","family":"Sauer","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biophysics Julius\u2010Maximilians\u2010Universit\u00e4t W\u00fcrzburg  Biozentrum W\u00fcrzburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00fcrgen","family":"Hemberger","sequence":"additional","affiliation":[{"name":"Institute for Biochemical Engineering and Analytics University of Applied Sciences Giessen Giessen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Jordan","sequence":"additional","affiliation":[{"name":"Merck Biopharma Biotech Process Sciences Fenil\u2010sur\u2010Corsier Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2019,2,27]]},"reference":[{"key":"e_1_2_7_1_2_1","doi-asserted-by":"publisher","DOI":"10.3390\/ph3010146"},{"key":"e_1_2_7_1_3_1","first-page":"1","article-title":"Delivering on the potential of biosimilar medicines: The role of functioning competitive markets introduction","author":"Aitken M.","year":"2016","journal-title":"IMS Health"},{"key":"e_1_2_7_1_4_1","doi-asserted-by":"publisher","DOI":"10.1155\/2013\/597282"},{"key":"e_1_2_7_1_5_1","doi-asserted-by":"publisher","DOI":"10.4161\/mabs.20996"},{"key":"e_1_2_7_1_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2015.09.014"},{"key":"e_1_2_7_1_7_1","unstructured":"Br\u00fchlmann D. 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