@article{ecae7396356f41859cdd452109a24fad,
title = "Generation of glycan-specific nanobodies",
abstract = "The development of antibodies that target specific glycan structures on cancer cells or human pathogens poses a significant challenge due to the immense complexity of naturally occurring glycans. Automated glycan assembly enables the production of structurally homogeneous glycans in amounts that are difficult to derive from natural sources. Nanobodies (Nbs) are the smallest antigen-binding domains of heavy-chain-only antibodies (hcAbs) found in camelids. To date, the development of glycan-specific Nbs using synthetic glycans has not been reported. Here, we use defined synthetic glycans for alpaca immunization to elicit glycan-specific hcAbs, and describe the identification, isolation, and production of a Nb specific for the tumor-associated carbohydrate antigen Globo-H. The Nb binds the terminal fucose of Globo-H and recognizes synthetic Globo-H in solution and native Globo-H on breast cancer cells with high specificity. These results demonstrate the potential of our approach for generating glycan-targeting Nbs to be used in biomedical and biotechnological applications.",
keywords = "nanobody, single domain antibody, heavy chain only antibody, glycan, glycoconjugate, immunization, globo H, tumor associated carbohydrate antigen, cancer, alpaca, active immunization",
author = "Khilji, {Sana Khan} and Felix Goerdeler and Kristin Frensemeier and David Warschkau and Jost L{\"u}hle and Zeinab Fandi and Falko Schirmeister and Chen, {Zhuo Angel} and Onur Turak and Alvaro Mallagaray and Stefan Boerno and Bernd Timmermann and Juri Rappsilber and Seeberger, {Peter H.} and Oren Moscovitz",
note = "Funding Information: The authors thank Prof. Phil Selenko for his careful and critical reading of the manuscript and Katrin Sellrie for her technical support. A.M. thanks Prof. Thomas Peters and the state of Schleswig-Holstein for supplying NMR infrastructure (European Funds for Regional Development, LPW-E/1.1.2/857). This work was supported by Deutsche Forschungsgemeinschaft (RTG2046) (F.G. and J.L.), the Wellcome Trust through core funding for The Wellcome Center for Cell Biology (203149), a Senior Research Fellowship (103139), and the Deutsche Forschungsgemeinschaft under Germany's Excellence Strategy, EXC 2008,390540038, UniSysCat. (J.R.), Alexander von Humboldt Foundation (O.M.), and the Max Planck Society. S.K.K. F.G. K.F. D.W. J.L. and Z.F. produced and characterized the Nbs described in this work. F.G. prepared glycoconjugates and performed glycan arrays. Z.A.C. and J.R. performed the MS analysis and database construction. Next-generation sequencing and cDNA library assembly were done by S.B. and B.T. Bioinformatic analysis was done by F.S. and O.T. A.M. characterized binding by nuclear magnetic resonance spectroscopy. F.G. and O.M. wrote the manuscript, with contributions from S.K.K. A.M. and P.H.S. to the final version. O.M. conceived the original idea and designed the concept and experiments. O.M. and P.H.S. supervised the project. O.M. and P.H.S. are co-founders of Tacalyx GmbH, where P.H.S. is a board member. O.M. and P.H.S. have a significant financial interest in the company and field a patent for the Nbs described in this manuscript (EP3799881A1). Funding Information: The authors thank Prof. Phil Selenko for his careful and critical reading of the manuscript and Katrin Sellrie for her technical support. A.M. thanks Prof. Thomas Peters and the state of Schleswig-Holstein for supplying NMR infrastructure (European Funds for Regional Development, LPW-E/1.1.2/857). This work was supported by Deutsche Forschungsgemeinschaft (RTG2046) (F.G. and J.L.), the Wellcome Trust through core funding for The Wellcome Center for Cell Biology (203149), a Senior Research Fellowship (103139), and the Deutsche Forschungsgemeinschaft under Germany's Excellence Strategy, EXC 2008,390540038, UniSysCat. (J.R.), Alexander von Humboldt Foundation (O.M.), and the Max Planck Society . Publisher Copyright: {\textcopyright} 2022 Elsevier Ltd",
year = "2022",
month = aug,
day = "18",
doi = "10.1016/j.chembiol.2022.05.007",
language = "English",
volume = "29",
pages = "1353--1361.e6",
journal = "Cell Chemical Biology",
issn = "2451-9456",
publisher = "Elsevier Inc.",
number = "8",
}