Characterization of molecular forms of probrain natriuretic peptide in human plasma
Characterization of molecular forms of probrain natriuretic peptide in human plasma
复制标题
DOI：
10.1016/s0009-8981(03)00240-7
复制
发表时间：
2003-08-01
影响因子：
5
通讯作者：
Sairenji, T
中科院分区：
文献类型：
--
作者：
Shimizu, H;Masuta, K;Sairenji, T
背景：血浆脑钠肽（BNP）的测定有助于心力衰竭的诊断和预后判断。然而，BNP的分子形式，特别是血液中的proBNP，特征很差。研究方法：为了研究proBNP在血液中的形式，开发了针对四种proBNP合成肽（1-13、22-27、28-40和4754）的抗体，并通过ELISA表征其对proBNP的反应性。利用这些抗体和BNP羧基端组氨酸特异性单克隆抗体，建立了proBNP的放射免疫分析法。通过凝胶过滤HPLC分析心力衰竭患者血浆中的BNP，然后进行BNP和proBNP的免疫测定。结果：4株抗proBNP合成肽抗体中有2株与患者血浆中的r-proBNP和BNP均呈阳性反应。凝胶过滤HPLC分析显示免疫反应性BNP的两个主峰（高MW BNP（36 kDa）和低MW BNP（4 kDa））。免疫分析表明，proBNP几乎全长包含在高分子量BNP组分中。结论：ProBNP分泌后在循环中比其成熟型BNP更稳定，提示proBNP测定的有效性。(C)2003 Elsevier B.V保留所有权利。
Background: Measurement of brain natriuretic peptide (BNP) in plasma is useful for the diagnosis and prognosis of heart failure. However, the molecular forms of BNP, especially proBNP in the blood, have been poorly characterized. Methods: To investigate the forms of proBNP in blood, antibodies against four proBNP synthetic peptides (1-13, 22-27, 28-40, and 4754) were developed and characterized for their reactivity to proBNP by ELISA. Using the antibodies and a monoclonal antibody specific to the carboxyl terminal histidine of BNP, a radioimmunoassay for proBNP was constructed. BNPs in plasma from heart failure patients were analyzed by gel-filtration HPLC followed by immunoassay for BNP and proBNP. Results: Two of four antibodies against proBNP synthetic peptides reacted with both r-proBNP and BNP in plasma from the patients. Gel-filtration HPLC analysis showed two major peaks of immunoreactive BNP (high MW BNP (36 kDa) and low MW BNP (4 kDa)). The immunoassay demonstrated that almost the full-length of proBNP was contained in the high MW BNP fractions. Conclusions: ProBNP was more stable than its mature form of BNP in circulation after secretion, and it suggested the usefulness of proBNP measurement. (C) 2003 Elsevier B.V All rights reserved.