The human rhabdomyosarcoma cell line TE671--Towards an innovative production platform for glycosylated biopharmaceuticals.
The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-translational modifications, in particular glycosylation, is a major issue. Glycans can influence ligand binding, serum half-life as well as biological activity or product immunogenicity. Aiming to establish a novel production platform for recombinant glycoproteins, the human TE671 cell line was investigated. Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. The cell line's ability to synthesize complex and highly sialylated N-glycan structures has been shown. Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems.
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.
Julia
J
Rosenlöcher
Laboratory of Biochemistry, Department of Life Sciences and Technology, Beuth University of Applied Sciences Berlin, Seestrasse 64, 13347 Berlin, Germany.
Constanze
C
Weilandt
Laboratory of Biochemistry, Department of Life Sciences and Technology, Beuth University of Applied Sciences Berlin, Seestrasse 64, 13347 Berlin, Germany.
Grit
G
Sandig
Laboratory of Biochemistry, Department of Life Sciences and Technology, Beuth University of Applied Sciences Berlin, Seestrasse 64, 13347 Berlin, Germany.
Stefan O
SO
Reinke
Institute of Laboratory Medicine, Clinical Chemistry and Pathobiochemistry, Charité - Universitätsmedizin Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
Véronique
V
Blanchard
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.
Stephan
S
Hinderlich
D001709:Biotechnology / Q000379:methods*; D002240:Carbohydrate Sequence; D045744:Cell Line, Tumor; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism*; D006031:Glycosylation; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism*
D006023:Glycoproteins; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor
Alpha-1-antitrypsin; Granulocyte–macrophage colony-stimulating factor; N-glycosylation; TE671; Therapeutic glycoprotein