A single-step, rapid, and versatile method for simultaneous detection of cell surface glycan profiles using fluorochrome-conjugated lectins

Author:

Torres Nicolás I123,Manselle-Cocco Montana N12,Perrotta Ramiro M12,Mahmoud Yamil D123,Salatino Mariana12,Mariño Karina V4,Rabinovich Gabriel A125ORCID

Affiliation:

1. Laboratorio de Glicomedicina , Programa de Glicociencias, , C1428, Ciudad de Buenos Aires, Argentina

2. Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) , Programa de Glicociencias, , C1428, Ciudad de Buenos Aires, Argentina

3. Universidad Argentina de la Empresa (UADE), Instituto de Tecnología (INTEC) , C1073, Ciudad de Buenos Aires, Argentina

4. Laboratorio de Glicómica Funcional y Molecular, Programa de Glicociencias, Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) , C1428, Ciudad de Buenos Aires, Argentina

5. Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires , C1428, Ciudad de Buenos Aires, Argentina

Abstract

Abstract Cell surface glycans play essential roles in diverse physiological and pathological processes and their assessment has important implications in biomedicine and biotechnology. Here we present a rapid, versatile, and single-step multicolor flow cytometry method for evaluation of cell surface glycan signatures using a panel of selected fluorochrome-conjugated lectins. This procedure allows simultaneous detection of cell surface glycans with a 10-fold reduction in the number of cells required compared with traditional multistep lectin staining methods. Interestingly, we used this one-step lectin array coupled with dimension reduction algorithms in a proof-of-concept application for discrimination among different tumor and immune cell populations. Moreover, this procedure was also able to unveil T-, B-, and myeloid cell subclusters exhibiting differential glycophenotypes. Thus, we report a rapid and versatile lectin cytometry method to simultaneously detect a particular repertoire of surface glycans on living cells that can be easily implemented in different laboratories and core facilities.

Funder

Ministerio de Ciencia, Tecnología e Innovación

Agencia Nacional de Promoción de la Investigación, el Desarrollo Tecnológico y la Innovación

Publisher

Oxford University Press (OUP)

Subject

Biochemistry

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