title: Recombinant glycoproteins: The impact of cell lines and culture conditions on the generation of protein species.
abstract: Glycosylation is the most complex post-translational modification. Thus, it contributes to versatile chemical compositions of proteins, leading to high amounts of protein species. The structural heterogeneity of glycoproteins was also described by the definition of glycoforms. We therefore introduced a new term called "glycoprotein species" to join the two concepts from different fields of biology. In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production.
  
  
  CONCLUSIONS
  Protein glycosylation is a major source for the huge amount of protein species. This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed.
authors: Laboratory of Biochemistry, Department of Life Sciences and Technology, Beuth University of Applied Sciences Berlin, Seestrasse 64, 13347 Berlin, Germany; Department of Biology, Chemistry and Pharmacy, Freie Universität Berlin, Takustrasse 3, 14195 Berlin, Germany.
authors: Julia
authors: J
authors: Rosenlöcher
authors: Laboratory of Biochemistry, Department of Life Sciences and Technology, Beuth University of Applied Sciences Berlin, Seestrasse 64, 13347 Berlin, Germany.
authors: Grit
authors: G
authors: Sandig
authors: Octapharma Biopharmaceuticals GmbH, Molecular Biochemistry Berlin, Octapharma R&D, Walther-Nernst-Strasse 3, 12489 Berlin, Germany.
authors: Christoph
authors: C
authors: Kannicht
authors: Institute of Laboratory Medicine, Clinical Chemistry and Pathobiochemistry, Charité - Universitätsmedizin Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
authors: Véronique
authors: V
authors: Blanchard
authors: Laboratory of Biochemistry, Department of Life Sciences and Technology, Beuth University of Applied Sciences Berlin, Seestrasse 64, 13347 Berlin, Germany.
authors: Stefan O
authors: SO
authors: Reinke
authors: Laboratory of Biochemistry, Department of Life Sciences and Technology, Beuth University of Applied Sciences Berlin, Seestrasse 64, 13347 Berlin, Germany. Electronic address: stephan.hinderlich@beuth-hochschule.de.
authors: Stephan
authors: S
authors: Hinderlich
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D000515:alpha 1-Antitrypsin / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D000515:alpha 1-Antitrypsin
keywords: Glycoform; Glycoprotein species; Microheterogeneity; N-glycosylation; Protein species; Recombinant glycoprotein