title: High molecular weight polyglycerol-based multivalent mannose conjugates.
abstract: We report the synthesis and characterization of multivalent mannose conjugates based on high molecular weight hyperbranched polyglycerols (HPG). A range of glycoconjugates were synthesized from high molecular weight HPGs (up to 493 kDa) and varying mannose units (22-303 per HPG). Hemagglutination assays using fresh human red blood cells and concanavalin A (Con A) showed that HPG-mannose conjugates exhibited a large enhancement in the relative potency of conjugates (as high as 40000) along with a significant increment in relative activity per sugar (up to 255). The size of the HPG scaffold and the number of mannose residues per HPG were all shown to influence the enhancement of binding interactions with Con A. Isothermal titration calorimetry (ITC) experiments confirmed the enhanced binding affinity and showed that both molecular size and ligand density play important roles. The enhancement in Con A binding to the high molecular weight HPG-mannose conjugates is due to a combination of inter- and intramolecular mannose binding. A few fold increments in the binding constant were obtained over mannose upon covalent attachment to HPG. The binding enhancement is due to the highly favorable entropic contribution to the multiple interactions of Con A to mannose residues on HPG. The high molecular weight HPG-mannose conjugates showed positive cooperativity in binding to Con A. Although carbohydrate density has less of an effect on functional valency of the conjugate compared to the molecular size, it determines the binding affinity.
authors.affiliation: Centre for Blood Research, Department of Pathology and Laboratory Medicine, Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada.
authors.forename: Jayachandran N
authors.initials: JN
authors.lastname: Kizhakkedathu
authors.forename: A Louise
authors.initials: AL
authors.lastname: Creagh
authors.forename: Rajesh A
authors.initials: RA
authors.lastname: Shenoi
authors.forename: Nicholas A A
authors.initials: NA
authors.lastname: Rossi
authors.forename: Donald E
authors.initials: DE
authors.lastname: Brooks
authors.forename: Timmy
authors.initials: T
authors.lastname: Chan
authors.forename: Jonathan
authors.initials: J
authors.lastname: Lam
authors.forename: Srinivasa R
authors.initials: SR
authors.lastname: Dandepally
authors.forename: Charles A
authors.initials: CA
authors.lastname: Haynes
mesh_terms: D003208:Concanavalin A / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism; D005990:Glycerol / Q000737:chemistry*; D006001:Glycoconjugates / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D006386:Hemagglutination Tests; D006801:Humans; D008358:Mannose / Q000737:chemistry*; D008970:Molecular Weight; D011108:Polymers / Q000737:chemistry*; D011485:Protein Binding
chemical_list: D006001:Glycoconjugates; D011108:Polymers; D003208:Concanavalin A; C043941:polyglycerol; D005990:Glycerol; D008358:Mannose