An antibody reactive to the Gly63-Lys68 epitope of NT-proBNP exhibits O-glycosylation-independent binding

Exp Mol Med. 2014 Sep 19;46(9):e114. doi: 10.1038/emm.2014.57.


The N-terminal fragment of prohormone brain natriuretic peptide (NT-proBNP) is a commonly used biomarker for the diagnosis of congestive heart failure, although its biological function is not well known. NT-proBNP exhibits heavy O-linked glycosylation, and it is quite difficult to develop an antibody that exhibits glycosylation-independent binding. We developed an antibody that binds to the recombinant NT-proBNP protein and its deglycosylated form with similar affinities in an enzyme immunoassay. The epitope was defined as Gly63-Lys68 based on mimetic peptide screening, site-directed mutagenesis and a competition assay with a peptide mimotope. The nearest O-glycosylation residues are Thr58 and Thr71; therefore, four amino acid residues intervene between the epitope and those residues in both directions. In conclusion, we report that an antibody reactive to Gly63-Lys68 of NT-proBNP exhibits O-glycosylation-independent binding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies / immunology*
  • Antigen-Antibody Reactions
  • Epitope Mapping
  • Epitopes / chemistry
  • Epitopes / genetics
  • Epitopes / immunology*
  • Glycosylation
  • HEK293 Cells
  • Heart Failure / immunology
  • Humans
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Natriuretic Peptide, Brain / chemistry
  • Natriuretic Peptide, Brain / genetics
  • Natriuretic Peptide, Brain / immunology*
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology*
  • Rabbits
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology


  • Antibodies
  • Epitopes
  • Peptide Fragments
  • Recombinant Fusion Proteins
  • pro-brain natriuretic peptide (1-76)
  • Natriuretic Peptide, Brain