title: Isolation and immobilization of influenza virus-specific N-SA-α-2,3-Gal receptors using magnetic nanoparticles coated with chitosan and Maackia amurensis lectin.
abstract: Avian influenza viruses preferentially bind to sialic acid alpha-2,3-galactose (N-SA-α-2,3-Gal) receptors on epithelial cells. Herein, we describe a procedure we have developed for isolation of N-SA-α-2,3-Gal receptors from porcine trachea using magnetic nanoparticles (NPs) coated with chitosan (NP-Ch) and functionalized with Maackia amurensis lectin (NP-lectin). Magnetic nanoparticles were coated with chitosan in a one-step co-precipitation, and then M. amurensis lectin was immobilized covalently using glutaraldehyde. Lectin coated nanoparticles were incubated with sialic acid enriched fraction of tracheal homogenate, and N-SA-α-2,3-Gal receptor was extracted under magnetic field in two cycles. The presence of 66.4 kDa protein was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The interaction of immobilized receptor (NP-Ch-R) with M. amurensis lectin (NP-Ch-R-L) was demonstrated by Fourier transform infrared spectrometry (FTIR) and thermogravimetric analysis (TGA).
authors.affiliation: Facultad de Ciencias Químicas, Grupo de Nanobiociencia de la Facultad de Ciencias Químicas de la Universidad Autónoma de Coahuila, Blvd. V. Carranza e Ing. José Cárdenas V. Col. República Oriente, C.P. 25280, Saltillo, Coahuila, Mexico.
authors.forename: Susana Araceli Carrizales
authors.initials: SA
authors.lastname: Alvarez
authors.forename: Anna
authors.initials: A
authors.lastname: Ilyina
authors.forename: Karla Monzerratt Gregorio
authors.initials: KM
authors.lastname: Jáuregui
authors.forename: José Luis Martínez
authors.initials: JL
authors.lastname: Hernández
authors.forename: Brenda Berenice Vazquez
authors.initials: BB
authors.lastname: Gutiérrez
authors.forename: Elda Patricia Segura
authors.initials: EP
authors.lastname: Ceniceros
authors.forename: Alejandro Zugasti
authors.initials: AZ
authors.lastname: Cruz
authors.forename: Hened Saade
authors.initials: HS
authors.lastname: Caballero
authors.forename: Raúl Guillermo López
authors.initials: RG
authors.lastname: Campos
mesh_terms: D000818:Animals; D048271:Chitosan / Q000737:chemistry*; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis; D018189:Immunomagnetic Separation / Q000379:methods; D029901:Maackia / Q000737:chemistry*; D060526:Magnetic Fields; D058185:Magnetite Nanoparticles / Q000737:chemistry* / Q000528:radiation effects / Q000648:ultrastructure; D009975:Orthomyxoviridae / Q000378:metabolism; D010316:Particle Size; D037121:Plant Lectins / Q000737:chemistry*; D011956:Receptors, Cell Surface / Q000302:isolation & purification* / Q000378:metabolism; D013552:Swine; D014132:Trachea / Q000737:chemistry*
chemical_list: D020099:Coated Materials, Biocompatible; D058185:Magnetite Nanoparticles; D037121:Plant Lectins; D011956:Receptors, Cell Surface; C039707:galactose receptor; C092488:sialic acid receptor; D048271:Chitosan