Comparative Antigenicity of Thiourea and Adipic Amide Linked Neoglycoconjugates Containing Modified Oligomannose Epitopes for the Carbohydrate-Specific anti-HIV Antibody 2G12

Bioconjug Chem. 2019 Jan 16;30(1):70-82. doi: 10.1021/acs.bioconjchem.8b00731. Epub 2018 Dec 28.


Novel neoglycoproteins containing oligomannosidic penta- and heptasaccharides as structural variants of oligomannose-type N-glycans found on human immunodeficiency virus type 1 gp120 have been prepared using different conjugation methods. Two series of synthetic ligands equipped with 3-aminopropyl spacer moieties and differing in the anomeric configuration of the reducing mannose residue were activated either as isothiocyanates or as adipic acid succinimidoyl esters and coupled to bovine serum albumin. Coupling efficiency for adipic acid connected neoglycoconjugates was better than for the thiourea-linked derivatives; the latter constructs, however, exhibited higher reactivity toward antibody 2G12, an HIV-neutralizing antibody with exquisite specificity for oligomannose-type glycans. 2G12 binding avidities for the conjugates, as determined by Bio-Layer Interferometry, were mostly higher for the β-linked ligands and, as expected, increased with the numbers of covalently linked glycans, leading to approximate KD values of 10 to 34 nM for optimized ligand-to-BSA ratios. A similar correlation was observed by enzyme-linked immunosorbent assays. In addition, dendrimer-type ligands presenting trimeric oligomannose epitopes were generated by conversion of the amino-spacer group into a terminal azide, followed by triazole formation using "click chemistry". The severe steric bulk of the ligands, however, led to poor efficiency in the coupling step and no increased antibody binding by the resulting neoglycoconjugates, indicating that the low degree of substitution and the spatial orientation of the oligomannose epitopes within these trimeric ligands are not conducive to multivalent 2G12 binding.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipates / chemistry*
  • Amides / chemistry
  • Carbohydrate Sequence
  • Click Chemistry
  • Dendrimers / chemistry
  • Epitopes / chemistry*
  • Glycoconjugates / chemical synthesis
  • Glycoconjugates / chemistry*
  • HIV Antibodies / chemistry
  • HIV Antibodies / immunology*
  • Mannose / chemistry*
  • Thiourea / chemistry*


  • Adipates
  • Amides
  • Dendrimers
  • Epitopes
  • Glycoconjugates
  • HIV Antibodies
  • adipic acid
  • Thiourea
  • Mannose