Assay for measurement of intact B-type natriuretic peptide prohormone in blood
Assay for measurement of intact B-type natriuretic peptide prohormone in blood
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DOI：
10.1373/clinchem.2005.061770
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发表时间：
2006-06-01
影响因子：
9.3
通讯作者：
Villard, Sylvie
中科院分区：
文献类型：
--
作者：
Giuliani, Isabelle;Rieunier, Francois;Villard, Sylvie
背景资料：据报道，B型利钠肽（BNP 1 -32）以及含有残基1-76的激素原的N-末端片段（NT-proBNP（1-76））（均为前体proBNP（1-108）的裂解产物）是心力衰竭预后和风险分层的有力标志物。然而，完整的前体也在血液中循环。已经开发了用于检测这些裂解产物的测定法，但是这些测定法中的大多数可能高估了裂解产物的浓度，因为它们也测量前体形式。因此，重要的是要发展一种免疫分析，专门测量proBNP（1-108）在plasma.Methods：经过精心设计的肽用于免疫小鼠，我们选择了一个特定的单克隆抗体（mAb Hinge 76），识别proBNP（1-108），一个表位的切割位点，只存在于前体形式。mAb Hinge 76以剂量依赖性方式识别重组proBNP（1-108），与重组NT-proBNP（1-76）或合成BNP 1 -32没有任何明显的交叉反应性。通过将mAb Hinge 76与针对BNP 1 -32的多克隆抗体结合，我们能够建立proBNP（1-108）特异性夹心免疫测定法，能够确认血液样本中proBNP（1-108）的存在。从50名健康人和170名充血性心力衰竭（CHF）患者的队列中，我们的测定能够区分健康个体和CHF患者（P < 0.005）。有趣的是，血浆proBNP（1-108）浓度与纽约心脏协会分级相关。结论：mAb Hinge 76是首个能够准确测定血浆proBNP（1 - 108）浓度的proBNP（1 - 108）特异性mAb。(c)2006年美国临床化学协会。
Background: B-Type natriuretic peptide (BNP1-32) as well as the N-terminal fragment of the prohormone containing residues 1-76 (NT-proBNP(1-76)), both cleavage products of the precursor proBNP(1-108), are reported to be powerful markers for prognosis and risk stratification of heart failure. However, the intact precursor also circulates in the bloodstream. Assays for the detection of these cleavage products have been developed, but most of these assays may overestimate the concentrations of the cleavage products because they also measure the precursor form. It is therefore important to develop an immunoassay that specifically measures solely proBNP(1-108) in plasma.Methods: After carefully designing the peptide used to immunize mice, we selected a specific monoclonal antibody (mAb Hinge76) that recognizes the cleavage site of proBNP(1-108), an epitope present only in the precursor form. mAb Hinge76 recognizes recombinant proBNP(1-108) in a dose-dependent manner, without any significant cross-reactivity with either recombinant NT-proBNP(1-76) or synthetic BNP1-32. By combining mAb Hinge76 with a polyclonal antibody directed against BNP1-32, we were able to set up a proBNP(1-108)-specific sandwich immunoassay able to confirm the presence of proBNP(1-108) in blood samples.Results: From a cohort of 50 healthy persons and 170 patients with congestive heart failure (CHF), our assay was able to differentiate healthy individuals from CHF patients (P < 0.005). Interestingly, plasma proBNP(1-108) concentrations were correlated with New York Heart Association classification. Moreover, a close relationship between proBNP(1-108) and BNP1-32 concentrations may exist, as a good correlation (r(2) = 0.89) was obtained when their respective concentrations were compared.Conclusion: mAb Hinge76 is the first proBNP(1-108)-specific mAb produced that allows accurate estimation of proBNP(1-108) concentrations in plasma. (c) 2006 American Association for Clinical Chemistry.