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KMID : 0620920140460090003
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Experimental & Molecular Medicine
2014 Volume.46 No. 9 p.3 ~ p.3
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An antibody reactive to the Gly63?Lys68 epitope of NT-proBNP exhibits O-glycosylation-independent binding
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Lee Yu-jean
Kim Hyo-Ri
Chung Jun-Ho
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Abstract
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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.
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