Nordic Bioscience, Fibrosis Biology and Biomarkers Herlev, Denmark.
Am J Transl Res. 2013 Apr 19;5(3):303-15. Print 2013.
The present study describes the assessment of true formation of type III collagen in different pathologies using a neo-epitope specific competitive Enzyme-linked immunosorbent assay (ELISA) towards the N-terminal propeptide of type III collagen (PRO-C3).
The monoclonal antibody was raised against the N-protease mediated cleavage site of the N-terminal propeptide of type III collagen and a competitive ELISA was developed using the selected antibody. The assay was evaluated in relation to neo-epitope specificity, technical performance, and as a marker for liver fibrosis and muscle mass using the rat carbon tetrachloride (CCl4) model and a study of immobilization induced muscle loss in humans, respectively.
The ELISA was neo-epitope specific, technically stable and can be assessed in serum and plasma samples. In the CCl4 liver fibrosis model it was observed that serum PRO-C3 were significantly elevated in rats with liver fibrosis as seen by histology (56% elevated in the highest quartile of total hepatic collagen compared to control rats, p<0.001) and correlated significantly to total hepatic collagen in the diseased rats (r=0.46, p<0.01) and not in control rats, suggesting the pathological origin of the epitope. Human plasma PRO-C3 correlated significantly to muscle mass at baseline (R(2)=0.44, p=0.036).
The developed neo-epitope specific serum ELISA for type III procollagen (PRO-C3) reflects true formation as it is specific for the propeptide cleaved off the intact collagen molecule. In a clinical and in a rodent study we showed that this marker was highly related to liver fibrosis and muscle mass.
本研究使用针对 III 型前胶原 N 端前肽（PRO-C3）的新型表位特异性竞争酶联免疫吸附试验（ELISA），描述不同病理状态下 III 型胶原真正形成的评估。
该单克隆抗体是针对 III 型前胶原 N 端前肽的 N 蛋白酶介导的切割位点产生的，并使用选定的抗体开发了竞争性 ELISA。该测定法与新型表位特异性、技术性能以及作为肝纤维化和肌肉质量标志物进行了评估，分别使用大鼠四氯化碳（CCl4）模型和人类固定诱导肌肉减少的研究。
ELISA 具有新型表位特异性，技术稳定，可在血清和血浆样本中进行评估。在 CCl4 肝纤维化模型中，通过组织学观察到纤维化大鼠的血清 PRO-C3 明显升高（与对照大鼠相比，总肝胶原的最高四分位数升高 56%，p<0.001），与患病大鼠的总肝胶原显著相关（r=0.46，p<0.01），与对照大鼠无关，表明表位的病理性起源。人血浆 PRO-C3 与基线时的肌肉质量显著相关（R（2）=0.44，p=0.036）。
开发的针对 III 型前胶原（PRO-C3）的新型表位特异性血清 ELISA 反映了真正的形成，因为它是针对从完整胶原分子上切割下来的前肽特异性的。在一项临床和啮齿动物研究中，我们表明该标志物与肝纤维化和肌肉质量高度相关。
