Glycosylation of Receptor Binding Domain of SARS-CoV-2 S-Protein Influences on Binding to Immobilized DNA Aptamers

Int J Mol Sci. 2022 Jan 5;23(1):557. doi: 10.3390/ijms23010557.


Nucleic acid aptamers specific to S-protein and its receptor binding domain (RBD) of SARS-CoV-2 (severe acute respiratory syndrome-related coronavirus 2) virions are of high interest as potential inhibitors of viral infection and recognizing elements in biosensors. Development of specific therapy and biosensors is complicated by an emergence of new viral strains bearing amino acid substitutions and probable differences in glycosylation sites. Here, we studied affinity of a set of aptamers to two Wuhan-type RBD of S-protein expressed in Chinese hamster ovary cell line and Pichia pastoris that differ in glycosylation patterns. The expression system for the RBD protein has significant effects, both on values of dissociation constants and relative efficacy of the aptamer binding. We propose glycosylation of the RBD as the main force for observed differences. Moreover, affinity of a several aptamers was affected by a site of biotinylation. Thus, the robustness of modified aptamers toward new virus variants should be carefully tested.

Keywords: SARS-CoV-2; aptamers; biolayer interferometry; coronavirus; dissociation constants; receptor-binding domain; sensor surface; spike glycoprotein.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Aptamers, Nucleotide / chemistry*
  • Aptamers, Nucleotide / metabolism*
  • Binding Sites
  • CHO Cells
  • Cricetulus
  • Glycosylation
  • Immobilized Nucleic Acids / metabolism*
  • Protein Binding
  • Protein Domains
  • Protein Interaction Domains and Motifs
  • Protein Processing, Post-Translational
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • SARS-CoV-2
  • Saccharomycetales / genetics
  • Spike Glycoprotein, Coronavirus / chemistry*
  • Spike Glycoprotein, Coronavirus / metabolism*


  • Aptamers, Nucleotide
  • Immobilized Nucleic Acids
  • Recombinant Proteins
  • Spike Glycoprotein, Coronavirus
  • spike protein, SARS-CoV-2

Supplementary concepts

  • Komagataella pastoris