Enzyme-linked immunosorbent serum assay specific for the 7S domain of Collagen Type IV (P4NP 7S): A marker related to the extracellular matrix remodeling during liver fibrogenesis
Enzyme-linked immunosorbent serum assay specific for the 7S domain of Collagen Type IV (P4NP 7S): A marker related to the extracellular matrix remodeling during liver fibrogenesis
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DOI：
10.1111/j.1872-034x.2011.00946.x
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发表时间：
2012-05-01
影响因子：
4.2
通讯作者：
Karsdal, Morten A.
中科院分区：
文献类型：
--
作者：
Leeming, Diana J.;Nielsen, Mette J.;Karsdal, Morten A.
目的：本文报道了一种新开发的IV型胶原7S结构域N端前肽（P4 NP 7S）竞争性酶联免疫吸附试验（ELISA）用于描述肝纤维化的能力。该测定应用了一种单克隆抗体特异性的PIVNP 7S表位100%同源的人，大鼠和小鼠species.Methods：单克隆抗体，提出了对选定的P4 NP 7S的特定序列。筛选抗体，并使用选定的抗体开发竞争性ELISA测定。该测定进行了评价，在有关的技术性能，并在两个临床前肝纤维化模型，胆管结扎模型（BDL）和四氯化碳模型（CCL 4）都在rats.Results：一个技术上强大的P4 NP 7S ELISA测定使用单克隆抗体。在BDL和CCL 4肝纤维化模型中，观察到肝纤维化大鼠中P4 NP 7S水平显著升高，如组织学所见（CCL 4：与对照组相比，总肝胶原最高四分位数升高283%，P = 0.001; BDL：与假手术组相比，第4周升高183%，P < 0.001），并与BDL大鼠肝脏IV型胶原表达量相关（r = 0.49，P < 0.05），而与假手术组相比（r = -0.12）。CCL 4处理组P4 NP 7S与总胶原含量呈显著正相关（P <0.0 0 1，r = 0.6 7），而对照组无相关性（r = 0.0 4）。该测定可改善纤维化定量。
Aim: The present study describes the ability of a newly developed N-terminal pro-peptides of type IV collagen 7S domain (P4NP 7S) competitive enzyme-linked immunosorbent assay (ELISA) for describing liver fibrosis. The assay applies a monoclonal antibody specific for a PIVNP 7S epitope 100% homologous in the human, rat, and mouse species.Methods: Monoclonal antibodies were raised against selected P4NP 7S specific sequences. Antibodies were screened and a competitive ELISA assay was developed using a selected antibody. The assay was evaluated in relation to technical performance, and in two preclinical liver fibrosis models; the bile duct ligation model (BDL) and the carbon tetrachloride model (CCL4) both performed in rats.Results: A technically robust P4NP 7S ELISA assay using a monoclonal antibody was produced. In the BDL and CCL4 liver fibrosis models it was observed that the P4NP 7S levels were significantly elevated in rat with liver fibrosis as seen by histology (CCL4: 283% elevated in the highest quartile of total hepatic collagen compared with controls, P = 0.001; BDL: 183% elevated at week 4 compared with sham, P < 0.001) and correlated to the amount of hepatic type IV collagen expression in BDL rats (r = 0.49, P < 0.05) in contrast to sham (r = -0.12). P4NP 7S also correlated to total collagen in CCL4 treated livers (P < 0.001, r = 0.67), however, not in controls (r = 0.04).Conclusions: This newly developed serum assay specific for P4NP 7S was highly related to liver fibrosis and correlated to extent of hepatic fibrosis. This assay may improve fibrosis quantification.