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Extraction and Immobilization of SA-α-2,6-Gal Receptors on Magnetic Nanoparticles to Study Receptor Stability and Interaction with Sambucus nigra Lectin

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Abstract

The interaction between influenza virus hemagglutinins and host cell with terminal sialic acid linked receptors, SA-α-2,6-Gal for human strains is important to obtain insights into this infectious disease. Sambucus nigra lectin has high affinity for SA-α-2,6-Gal receptors. The goals of this work were: to extract the SA-α-2,6-Gal receptors from porcine airways; to perform receptors immobilization and study their storage stability; and to determine some parameters of interaction between the receptor and S. nigra lectin. The receptor isolation was monitored by means of bound sialic acid (BSAc) detection. A major band of protein at 66.7 kDa was clearly visible in SDS-PAGE assay. Eighty-one percent of isolated glycoproteins were immobilized on magnetic nanoparticles. The kinetics of BSAc storage stability at 4 °C was approximated as the first order reaction with kinetic constant and half-life estimated as 0.062 day−1 and 11.2 days, respectively. The dissociation constant (K d) calculated from Scatchard's plot was 2.47 × 10−7 M, and the receptor concentration was equal to 7.92 × 10−5 M. Procedure for N-SA-α-2,6-Gal -receptors extraction based on their affinity to S. nigra lectin with magnetic nanoparticles, and their immobilization in active form, was not described previously, and may have wide application in designing biosensors or virus removal from areas or contaminated samples.

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Acknowledgements

The authors are grateful to Julieta Sánchez (CIQA) for her technical assistance in assay related to nanoparticles characterization. We thank CONACYT for Ph.D. thesis scholarship.

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Authors and Affiliations

  1. Grupo de Nanobiociencia de la Facultad de Ciencias Químicas, Universidad Autónoma de Coahuila, Boulevard V. Carranza y José Cárdenas Valdés, 25280, Saltillo, Coahuila, Mexico

    Karla M. Gregorio-Jauregui, Susana A. Carrizalez-Alvarez, Jorge E. Rivera-Salinas, José L. Martinez, Elda P. Segura & Anna Ilyina

  2. Departamento de Procesos de Polimerización, Centro de Investigación en Química Aplicada, Boulevard Enrique Reyna No. 140, 25294, Saltillo, Coahuuila, Mexico

    Hened Saade & Raúl G. López

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  1. Karla M. Gregorio-Jauregui
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Correspondence to Anna Ilyina.

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Gregorio-Jauregui, K.M., Carrizalez-Alvarez, S.A., Rivera-Salinas, J.E. et al. Extraction and Immobilization of SA-α-2,6-Gal Receptors on Magnetic Nanoparticles to Study Receptor Stability and Interaction with Sambucus nigra Lectin. Appl Biochem Biotechnol 172, 3721–3735 (2014). https://doi.org/10.1007/s12010-014-0801-x

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