title: Ring-opening metathesis polymerization-derived, lectin-functionalized monolithic supports for affinity separation of glycoproteins.
abstract: Lectin-functionalized monolithic columns were prepared within polyether ether ketone (PEEK) columns (150 × 4.6 mm id) via transition metal-catalyzed ring-opening metathesis polymerization of norborn-2-ene (NBE) and trimethylolpropane-tris(5-norbornene-2-carboxylate) (CL) using the first-generation Grubbs initiator RuCl2 (PCy3 )2 (CHPh) (1, Cy = cyclohexyl) in the presence of a macro- and microporogen, i.e. of 2-propanol and toluene. Postsynthesis functionalization was accomplished via in situ grafting of 2,5-dioxopyrrolidin-1-yl-bicyclo[2.2.1]hept-5-ene-2-carboxylate to the surface of the monoliths followed by reaction with α,ω-diamino-poly(ethyleneglycol). The pore structure of the poly(ethyleneglycol)- derivatized monoliths was investigated by electron microscopy and inverse-size exclusion chromatography, respectively. The amino-poly(ethyleneglycol) functionalized monolithic columns were then successfully used for the immobilization of lectin from Lens culinaris hemagglutinin. The thus prepared lectin-functionalized monoliths were applied to the affinity chromatography-based purification of glucose oxidase. The binding capacity of Lens culinaris hemagglutinin-immobilized monolithic column for glucose oxidase was found to be 2.2 mg/column.
authors.affiliation: Lehrstuhl für Makromolekulare Stoffe und Faserchemie, Institut für Polymerchemie, Universität Stuttgart, Stuttgart, Germany.
authors.forename: Rajendar
authors.initials: R
authors.lastname: Bandari
authors.forename: Jürgen
authors.initials: J
authors.lastname: Kuballa
authors.forename: Michael R
authors.initials: MR
authors.lastname: Buchmeiser
mesh_terms: D001577:Benzophenones; D002384:Catalysis; D002846:Chromatography, Affinity; D005949:Glucose Oxidase / Q000032:analysis / Q000737:chemistry; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry*; D007659:Ketones / Q000737:chemistry; D037102:Lectins / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning; D015394:Molecular Structure; D011092:Polyethylene Glycols / Q000737:chemistry; D058105:Polymerization; D011108:Polymers; D011485:Protein Binding; D015203:Reproducibility of Results
chemical_list: D001577:Benzophenones; D006023:Glycoproteins; D007659:Ketones; D037102:Lectins; D011108:Polymers; C063834:polyetheretherketone; D011092:Polyethylene Glycols; D005949:Glucose Oxidase