A combined computational-experimental approach to define the structural origin of antibody recognition of sialyl-Tn, a tumor-associated carbohydrate antigen.

Autor: Amon R; Department of Cell Research and Immunology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, 69978, Israel., Grant OC; Complex Carbohydrate Research Center, University of Georgia, Athens, 30606, GA, USA., Leviatan Ben-Arye S; Department of Cell Research and Immunology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, 69978, Israel., Makeneni S; Complex Carbohydrate Research Center, University of Georgia, Athens, 30606, GA, USA., Nivedha AK; Complex Carbohydrate Research Center, University of Georgia, Athens, 30606, GA, USA., Marshanski T; Department of Cell Research and Immunology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, 69978, Israel., Norn C; Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, 76100, Israel., Yu H; Department of Chemistry, University of California-Davis, Davis, CA, USA., Glushka JN; Complex Carbohydrate Research Center, University of Georgia, Athens, 30606, GA, USA., Fleishman SJ; Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, 76100, Israel., Chen X; Department of Chemistry, University of California-Davis, Davis, CA, USA., Woods RJ; Complex Carbohydrate Research Center, University of Georgia, Athens, 30606, GA, USA. rwoods@ccrc.uga.edu., Padler-Karavani V; Department of Cell Research and Immunology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, 69978, Israel. vkaravani@post.tau.ac.il.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2018 Jul 17; Vol. 8 (1), pp. 10786. Date of Electronic Publication: 2018 Jul 17.
DOI: 10.1038/s41598-018-29209-9
Abstrakt: Anti-carbohydrate monoclonal antibodies (mAbs) hold great promise as cancer therapeutics and diagnostics. However, their specificity can be mixed, and detailed characterization is problematic, because antibody-glycan complexes are challenging to crystallize. Here, we developed a generalizable approach employing high-throughput techniques for characterizing the structure and specificity of such mAbs, and applied it to the mAb TKH2 developed against the tumor-associated carbohydrate antigen sialyl-Tn (STn). The mAb specificity was defined by apparent K D values determined by quantitative glycan microarray screening. Key residues in the antibody combining site were identified by site-directed mutagenesis, and the glycan-antigen contact surface was defined using saturation transfer difference NMR (STD-NMR). These features were then employed as metrics for selecting the optimal 3D-model of the antibody-glycan complex, out of thousands plausible options generated by automated docking and molecular dynamics simulation. STn-specificity was further validated by computationally screening of the selected antibody 3D-model against the human sialyl-Tn-glycome. This computational-experimental approach would allow rational design of potent antibodies targeting carbohydrates.
Databáze: MEDLINE
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