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Versatile strategy for the synthesis of biotin-labelled glycans, their immobilization to establish a bioactive surface and interaction studies with a lectin on a biochip

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Abstract

The emerging role of glycans as versatile biochemical signals in diverse aspects of cellular sociology calls for establishment of sensitive methods to monitor carbohydrate recognition by receptors such as lectins. Most of these techniques involve the immobilization of one of the binding partners on a surface, e.g. atomic force microscopy, glycan array and Surface Plasmon Resonance (SPR), hereby simulating cell surface presentation. Here, we report the synthesis of fluorescent glycoconjugates, with a functionalization strategy which avoids the frequently occurring ring opening at the reducing end for further immobilization on a surface or derivatization with biotin. In order to improve the versatility of these derivatized glycans for biological studies, a new approach for the synthesis of biotinylated and fluorescent glycans has also been realized. Finally, to illustrate their usefulness the neoglycoconjugates were immobilized on different surfaces, and the interaction analysis with a model lectin, the toxin from mistletoe, proved them to act as potent ligands, underscoring the merit of the presented synthetic approach.

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Acknowledgement

This work was supported by a Research Project of the MEC (Ministerio de Educación y Ciencia de España, CTQ2006-09052/BQU), a European Project (FP-62003-NMP-SMF-3, proposal 011774-2), an EC Marie Curie Research Training Network grant (MRTN-CT-2005-019561), the research initiative LMUexcellent and one of the authors (F. Javier Muñoz) thanks the MEC for a Ph.D. grant (MEC-FPU predoctoral fellowship AP2003-4820).

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Authors and Affiliations

  1. Grupo de Biotransformaciones, Departamento de Química Orgánica y Farmacéutica, Facultad de Farmacia, Universidad Complutense de Madrid (UCM), Pz/ Ramón y Cajal s/n., 28040, Madrid, Spain

    F. Javier Muñoz, José V. Sinisterra & María J. Hernáiz

  2. Departamento de Química Orgánica, Facultad de Químicas, Universidad Autónoma (UAM), 28049, Cantoblanco, Madrid, Spain

    Ángel Rumbero

  3. Servicio de Biotransformaciones Industriales, Parque Científico de Madrid, Tres Cantos, 28760, Madrid, Spain

    José V. Sinisterra

  4. Departamento de Ciencia de Proteínas, Centro de Investigaciones Biológicas, CIB-CSIC, Ramiro de Maeztu 9, 28040, Madrid, Spain

    J. Ignacio Santos & Jesús Jiménez-Barbero

  5. Institut für Physiologische Chemie, Tierärztliche Fakultät, Ludwig-Maximilians-Universität München, Veterinärstr. 13, 80539, München, Germany

    Sabine André & Hans-J. Gabius

  6. Servicio de Interacciones Biomoleculares, Parque Científico de Madrid, Facultad de Farmacia, Universidad Complutense, Pz/ Ramón y Cajal s/n., 28040, Madrid, Spain

    María J. Hernáiz

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  1. F. Javier Muñoz
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  2. Ángel Rumbero
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  3. José V. Sinisterra
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  4. J. Ignacio Santos
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  5. Sabine André
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  6. Hans-J. Gabius
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  7. Jesús Jiménez-Barbero
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  8. María J. Hernáiz
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Correspondence to María J. Hernáiz.

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Muñoz, F.J., Rumbero, Á., Sinisterra, J.V. et al. Versatile strategy for the synthesis of biotin-labelled glycans, their immobilization to establish a bioactive surface and interaction studies with a lectin on a biochip. Glycoconj J 25, 633–646 (2008). https://doi.org/10.1007/s10719-008-9115-y

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