An antibody reactive to the Gly63-Lys68 epitope of NT-proBNP exhibits O-glycosylation-independent binding
Experimental & Molecular Medicine
; : e114-2014.
Article
in En
| WPRIM
| ID: wpr-50917
Responsible library:
WPRO
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.
Full text:
1
Database:
WPRIM
Main subject:
Peptide Fragments
/
Glycosylation
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Recombinant Fusion Proteins
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Amino Acid Sequence
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Epitope Mapping
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Natriuretic Peptide, Brain
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HEK293 Cells
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Heart Failure
Limits:
Animals
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Humans
Language:
En
Journal:
Experimental & Molecular Medicine
Year:
2014
Document type:
Article