An antibody reactive to the Gly63-Lys68 epitope of NT-proBNP exhibits O-glycosylation-independent binding.
- Author:
Yujean LEE
1
;
Hyori KIM
;
Junho CHUNG
Author Information
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- MeSH: Amino Acid Sequence; Animals; Antibodies/*immunology; Antigen-Antibody Reactions; Epitope Mapping; Epitopes/chemistry/genetics/*immunology; Glycosylation; HEK293 Cells; Heart Failure/immunology; Humans; Molecular Sequence Data; Mutagenesis, Site-Directed; Natriuretic Peptide, Brain/chemistry/genetics/*immunology; Peptide Fragments/chemistry/genetics/*immunology; Rabbits; Recombinant Fusion Proteins/chemistry/genetics/immunology
- From:Experimental & Molecular Medicine 2014;46(9):e114-
- CountryRepublic of Korea
- Language:English
- Abstract: 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.
