title: Selective capture of glycoproteins using lectin-modified nanoporous gold monolith.
abstract: The surface of nanoporous gold (np-Au) monoliths was modified via a flow method with the lectin Concanavalin A (Con A) to develop a substrate for separation and extraction of glycoproteins. Self-assembled monolayers (SAMs) of α-lipoic acid (LA) on the np-Au monoliths were prepared followed by activation of the terminal carboxyl groups to create amine reactive esters that were utilized in the immobilization of Con A. Thermogravimetric analysis (TGA) was used to determine the surface coverages of LA and Con A on np-Au monoliths which were found to be 1.31×10(18) and 1.85×10(15)moleculesm(-2), respectively. An in situ solution depletion method was developed that enabled surface coverage characterization without damaging the substrate and suggesting the possibility of regeneration. Using this method, the surface coverages of LA and Con A were found to be 0.989×10(18) and 1.32×10(15)moleculesm(-2), respectively. The selectivity of the Con A-modified np-Au monolith for the high mannose-containing glycoprotein ovalbumin (OVA) versus negative control non-glycosylated bovine serum albumin (BSA) was demonstrated by the difference in the ratio of the captured molecules to the immobilized Con A molecules, with OVA:Con A=2.3 and BSA:Con A=0.33. Extraction of OVA from a 1:3 mole ratio mixture with BSA was demonstrated by the greater amount of depletion of OVA concentration during the circulation with the developed substrate. A significant amount of captured OVA was eluted using α-methyl mannopyranoside as a competitive ligand. This work is motivated by the need to develop new materials for chromatographic separation and extraction substrates for use in preparative and analytical procedures in glycomics.
authors.affiliation: Department of Chemistry and Biochemistry, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121 USA; Center for Nanoscience, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121, USA.
authors.forename: Allan J
authors.initials: AJ
authors.lastname: Alla
authors.affiliation: Department of Chemistry and Biochemistry, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121 USA; Center for Nanoscience, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121, USA.
authors.forename: Felipe B
authors.initials: FB
authors.lastname: D' Andrea
authors.affiliation: Department of Chemistry and Biochemistry, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121 USA; Center for Nanoscience, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121, USA.
authors.forename: Jay K
authors.initials: JK
authors.lastname: Bhattarai
authors.affiliation: Department of Chemistry and Biochemistry, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121 USA; Center for Nanoscience, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121, USA.
authors.forename: Jared A
authors.initials: JA
authors.lastname: Cooper
authors.affiliation: Department of Chemistry and Biochemistry, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121 USA; Center for Nanoscience, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121, USA.
authors.forename: Yih Horng
authors.initials: YH
authors.lastname: Tan
authors.affiliation: Department of Chemistry and Biochemistry, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121 USA.
authors.forename: Alexei V
authors.initials: AV
authors.lastname: Demchenko
authors.affiliation: Department of Chemistry and Biochemistry, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121 USA; Center for Nanoscience, One University Boulevard, University of Missouri-St. Louis, Saint Louis, MO 63121, USA. Electronic address: kstine@umsl.edu.
authors.forename: Keith J
authors.initials: KJ
authors.lastname: Stine
mesh_terms: D000818:Animals; D002417:Cattle; D003208:Concanavalin A / Q000737:chemistry; D054794:Glycomics / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification*; D006046:Gold / Q000737:chemistry*; D037102:Lectins / Q000737:chemistry*; D010047:Ovalbumin / Q000737:chemistry; D008063:Thioctic Acid / Q000737:chemistry
chemical_list: D006023:Glycoproteins; D037102:Lectins; D003208:Concanavalin A; D008063:Thioctic Acid; D006046:Gold; D010047:Ovalbumin
keywords: Chromatography; Glycomics; Glycoprotein; Lectin; Monolith; Nanoporous gold