title: Application of Lectin Array Technology for Biobetter Characterization: Its Correlation with FcγRIII Binding and ADCC.
abstract: Lectin microarray technology was applied to compare the glycosylation pattern of the monoclonal antibody MB311 expressed in SP2.0 cells to an antibody-dependent cellular cytotoxic effector function (ADCC)-optimized variant (MB314). MB314 was generated by a plant expression system that uses genetically modified moss protoplasts (Physcomitrella patens) to generate a de-fucosylated version of MB311. In contrast to MB311, no or very low interactions of MB314 with lectinsAspergillus oryzael-fucose (AOL),Pisum sativumagglutinin (PSA),Lens culinarisagglutinin (LCA), andAleuria aurantialectin (AAL) were observed. These lectins are specific for mono-/biantennary N-glycans containing a core fucose residue. Importantly, this fucose indicative lectin-binding pattern correlated with increased MB314 binding to CD16 (FcγRIII; receptor for the constant region of an antibody)-whose affinity is mediated through core fucosylation-and stronger ADCC. In summary, these results demonstrate that lectin microarrays are useful orthogonal methods during antibody development and for characterization.
authors.affiliation: Vela Labs GmbH, Brunner Str. 69/ Obj. 3, 1230 Vienna, Austria. m.roucka@vela-labs.at.
authors.forename: Markus
authors.initials: M
authors.lastname: Roucka
authors.affiliation: Vela Labs GmbH, Brunner Str. 69/ Obj. 3, 1230 Vienna, Austria. k.zimmermann@vela-labs.at.
authors.forename: Klaus
authors.initials: K
authors.lastname: Zimmermann
authors.affiliation: Vela Labs GmbH, Brunner Str. 69/ Obj. 3, 1230 Vienna, Austria. m.fido@vela-labs.at.
authors.forename: Markus
authors.initials: M
authors.lastname: Fido
authors.affiliation: Vela Labs GmbH, Brunner Str. 69/ Obj. 3, 1230 Vienna, Austria. anechansky@jhlbiotech.com.|JHL Biotech, Zhubei City, Hsinchu County 302, Taiwan. anechansky@jhlbiotech.com.
authors.forename: Andreas
authors.initials: A
authors.lastname: Nechansky
keywords: ADCC; GlycoStation; LecChip; anti-Lewis Y mAb; fucosylation; glyco-profiling; lectin carbohydrates; lectin microarray