title: Oligosaccharides and peptide displayed on an amphiphilic polymer enable solid phase assay of hapten specific antibodies.
abstract: Copovidone, a copolymer of vinyl acetate and N-vinyl-2-pyrrolidone, was synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization, and after deacetylation the polymer was functionalized by introduction of amino, azide, and alkyne pendant groups to allow attachment of glycans and peptide. Candida albicans β-mannan trisaccharides 1 and 2 and M. tuberculosis arabinan hexasaccharide 3 with appropriate tethers were conjugated to the polymers by squarate or click chemistry. C. albicans T-cell peptide 4 bearing a C-terminal ε-azidolysine was also conjugated to copovidone by click chemistry. The resulting conjugates provide convenient non-protein-based antigens that are readily adsorbed on ELISA plates, and display excellent characteristics for assay of antibody binding to the haptenic group of interest. Copovidone and BSA glycoconjugates exhibited similar adsorption characteristics when used to coat ELISA plates, and both conjugates were optimal when used as coating solutions at low nanogram/mL concentrations. Provided that the copovidone conjugated glycan is stable to acid, assay plates can be easily processed for reuse at least three times without detectable variation or degradation in ELISA readout.
authors.affiliation: Alberta Glycomics Centre, Department of Chemistry, University of Alberta , Edmonton, Alberta, Canada T6G 2G2.
authors.forename: David R
authors.initials: DR
authors.lastname: Bundle
authors.forename: Pui-Hang
authors.initials: PH
authors.lastname: Tam
authors.forename: Huu-Anh
authors.initials: HA
authors.lastname: Tran
authors.forename: Eugenia
authors.initials: E
authors.lastname: Paszkiewicz
authors.forename: Jonathan
authors.initials: J
authors.lastname: Cartmell
authors.forename: Joanna M
authors.initials: JM
authors.lastname: Sadowska
authors.forename: Susmita
authors.initials: S
authors.lastname: Sarkar
authors.forename: Maju
authors.initials: M
authors.lastname: Joe
authors.forename: Pavel I
authors.initials: PI
authors.lastname: Kitov
mesh_terms: D000327:Adsorption; D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D000918:Antibody Specificity; D002176:Candida albicans / Q000737:chemistry / Q000276:immunology; D057930:Click Chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D017743:Equipment Reuse; D006241:Haptens / Q000737:chemistry / Q000276:immunology*; D008968:Molecular Conformation; D009169:Mycobacterium tuberculosis / Q000737:chemistry / Q000276:immunology; D009844:Oligosaccharides / Q000737:chemistry / Q000276:immunology*; D010455:Peptides / Q000737:chemistry / Q000276:immunology*; D011108:Polymers / Q000737:chemistry; D011759:Pyrrolidines / Q000737:chemistry / Q000276:immunology*; D013501:Surface-Active Agents / Q000737:chemistry*; D014753:Vinyl Compounds / Q000737:chemistry / Q000276:immunology*
chemical_list: D000906:Antibodies; D006241:Haptens; D009844:Oligosaccharides; D010455:Peptides; D011108:Polymers; D011759:Pyrrolidines; D013501:Surface-Active Agents; D014753:Vinyl Compounds; C402301:poly(vinylpyrrolidone-co-vinyl-acetate)