title: Temperature-responsive size-exclusion chromatography using poly(N-isopropylacrylamide) grafted silica.
abstract: Silica-based packing materials induce non-specific interactions with proteins in aqueous media because of the nature of their surface, mainly silanol groups. Therefore, the silica surface has to be modified in order to be used as stationary phase for the High Performance Size-Exclusion Chromatography (HPSEC) of proteins. For this purpose, porous silica beads were coated with hydrophilic polymer gels (dextrans of different molecular weights) carrying a calculated amount of diethyl-aminoethyl groups (DEAE). Actually, as shown by HPSEC, these dextran modified supports minimize non-specific adsorption for proteins and pullulans in aqueous solution. Then, in order to change the pore size in response to temperature, temperature responsive polymer of poly(N-isopropylacrylamide) (PIPAAm) was introduced into the surface of dextran-DEAE on porous silica beads. The structure of these supports before and after modification was alternately studied by Scanning Electronic Microscopy (SEM) and Scanning Force Microscopy (SFM). An adsorption of radiolabelled albumin was performed to complete our study. Silica modifications by dextran-DEAE and PIPAAm improve the neutrality of the support and minimize the non-specific interactions between the solid support and proteins in solution. At low temperature, the support having PIPAAm exhibits a high resolution domain in HPSEC and finally permits a better resolution of proteins and pullulans. At higher temperature, hydrophobic properties of PIPAAm produce interactions with some proteins and trigger off a slight delay of their elution time.
authors.affiliation: Laboratoire de recherche sur les macromolécules, CNRS URA 502, Université Paris Nord, Institut Galilée, Villetaneuse, France.
authors.forename: H
authors.initials: H
authors.lastname: Lakhiari
authors.forename: T
authors.initials: T
authors.lastname: Okano
authors.forename: N
authors.initials: N
authors.lastname: Nurdin
authors.forename: C
authors.initials: C
authors.lastname: Luthi
authors.forename: P
authors.initials: P
authors.lastname: Descouts
authors.forename: D
authors.initials: D
authors.lastname: Muller
authors.forename: J
authors.initials: J
authors.lastname: Jozefonvicz
mesh_terms: D000180:Acrylic Resins / Q000138:chemical synthesis / Q000737:chemistry; D000327:Adsorption; D002850:Chromatography, Gel / Q000295:instrumentation* / Q000379:methods*; D006801:Humans; D018625:Microscopy, Atomic Force; D008855:Microscopy, Electron, Scanning; D009994:Osmolar Concentration; D012711:Serum Albumin, Radio-Iodinated / Q000378:metabolism; D012822:Silicon Dioxide / Q000138:chemical synthesis / Q000737:chemistry; D013499:Surface Properties; D013696:Temperature
chemical_list: D000180:Acrylic Resins; D012711:Serum Albumin, Radio-Iodinated; C052970:poly-N-isopropylacrylamide; D012822:Silicon Dioxide