Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity
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Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity. / Calow, Jenny; Behrens, Anna-Janina; Mader, Sonja; Bockau, Ulrike; Struwe, Weston B; Harvey, David J; Cormann, Kai U; Nowaczyk, Marc M; Loser, Karin; Schinor, Daniel; Hartmann, Marcus W W; Crispin, Max.
in: MABS-AUSTIN, Jahrgang 8, Nr. 8, 30.10.2016, S. 1498-1511.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity
AU - Calow, Jenny
AU - Behrens, Anna-Janina
AU - Mader, Sonja
AU - Bockau, Ulrike
AU - Struwe, Weston B
AU - Harvey, David J
AU - Cormann, Kai U
AU - Nowaczyk, Marc M
AU - Loser, Karin
AU - Schinor, Daniel
AU - Hartmann, Marcus W W
AU - Crispin, Max
PY - 2016/10/30
Y1 - 2016/10/30
N2 - Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. The resulting antibody demonstrated enhanced antibody-dependent cell-mediated cytotoxicity, which we attribute to unusual N-linked glycosylation. Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. In addition, terminal glucose structures were identified that match biosynthetic precursors of human glycosylation. Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity.
AB - Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. The resulting antibody demonstrated enhanced antibody-dependent cell-mediated cytotoxicity, which we attribute to unusual N-linked glycosylation. Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. In addition, terminal glucose structures were identified that match biosynthetic precursors of human glycosylation. Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity.
KW - Journal Article
U2 - 10.1080/19420862.2016.1228504
DO - 10.1080/19420862.2016.1228504
M3 - SCORING: Journal article
C2 - 27594301
VL - 8
SP - 1498
EP - 1511
JO - MABS-AUSTIN
JF - MABS-AUSTIN
SN - 1942-0862
IS - 8
ER -
