Development of fragment-specific osteopontin antibodies and ELISA for quantification in human metastatic breast cancer.
Development of fragment-specific osteopontin antibodies and ELISA for quantification in human metastatic breast cancer.
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碎片特异性骨桥蛋白抗体和ELISA的开发用于人类转移性乳腺癌。
DOI：
10.1186/1471-2407-8-38
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发表时间：
2008-01-31
期刊：
影响因子：
3.8
通讯作者：
Liaw, Lucy
中科院分区：
文献类型：
--
作者：
Plumer, Alicia;Duan, Hongyi;Subramaniam, Sripriya;Lucas, F. Lee;Miesfeldt, Susan;Ng, Ah-Kau;Liaw, Lucy
骨桥蛋白 (OPN) 与人类癌症相关，循环血液 OPN 可能在临床肿瘤学中具有诊断或预后价值。为了评估 OPN 作为癌症生物标志物，我们生成并表征了五种针对人全长 OPN (fl-OPN) 的新型小鼠单克隆抗体。四种抗体（2C5、2F10、2H9 和 2E11）识别的表位映射到 N 端 OPN (aa1-166)；一个 (1F11) 映射到 C 端 OPN (aa167-314)。这些抗体通过 ELISA 和免疫印迹识别重组和天然 OPN，与人和小鼠 OPN 发生交叉反应。其中两种新型抗体（2F10 和 1F11）用于开发 fl-OPN 的定量酶联免疫吸附测定 (ELISA)。与市售 ELISA 相比，我们的检测方法在测量 fl-OPN 标准品方面具有较高的准确性和灵敏度。具体来说，我们的 ELISA 具有 0.078 ng/ml-10 ng/ml 之间的线性剂量响应，灵敏度为 13.9 pg/ml。我们利用这种测定法对健康志愿者血浆中的 fl-OPN 进行定量，并与转移性乳腺癌患者进行比较。健康志愿者的平均循环血浆 fl-OPN 为 1.2 ng/ml，而转移性乳腺癌患者的平均循环血浆 fl-OPN 为 4.76 ng/ml (p = 0.0042)。尽管癌症患者中 fl-OPN 的增加与之前的研究一致，但所有现有 fl-OPN ELISA 的测量数量差异很大。由于 OPN 是一种复杂分子，具有可变剪接、翻译后修饰、细胞外蛋白水解修饰和参与蛋白质复合物等多样性，因此我们建议进一步了解多种 OPN 物种的特定亚型识别对于 OPN 生物标志物效用的未来研究至关重要。
Osteopontin (OPN) is associated with human cancers, and circulating blood OPN may have diagnostic or prognostic value in clinical oncology. To evaluate OPN as a cancer biomarker, we generated and characterized five novel mouse monoclonal antibodies against the human full-length OPN (fl-OPN). Epitopes recognized by four antibodies (2C5, 2F10, 2H9, and 2E11) map to N-terminal OPN (aa1-166); one (1F11) maps to C-terminal OPN (aa167-314). These antibodies recognize recombinant and native OPN by ELISA and immunoblot, cross reacting with human and mouse OPN. Two of these novel antibodies (2F10 and 1F11) were used to develop a quantitative enzyme linked immunosorbent assay (ELISA) for fl-OPN. In comparison with commercially available ELISAs, our assay had high accuracy in measuring fl-OPN standards, and high sensitivity. Specifically, our ELISA has a linear dose response between 0.078 ng/ml-10 ng/ml, with a sensitivity of 13.9 pg/ml. We utilized this assay to quantify fl-OPN in the plasma of healthy volunteers in comparison with patients with metastatic breast cancer. The average circulating plasma fl-OPN in healthy volunteers was 1.2 ng/ml, compared to 4.76 ng/ml in patients with metastatic breast cancer (p = 0.0042). Although the increase in fl-OPN in cancer patients is consistent with previous studies, the measured quantity varied greatly between all existing fl-OPN ELISAs. Because OPN is a complex molecule with diversity from alternative splicing, post-translational modification, extracellular proteolytic modification, and participation in protein complexes, we suggest that further understanding of specific isoform recognition of multiple OPN species is essential for future studies of OPN biomarker utility.
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影响因子：
11.5
作者：
Bramwell, Vivien H. C.;Doig, Gordon S.;Chambers, Ann F.
通讯作者：
Chambers, Ann F.
影响因子：
3.3
作者：
BROWN, LF;BERSE, B;SENGER, DR
通讯作者：
SENGER, DR
影响因子：
6.9
作者：
Gao, YA;Agnihotri, R;Liaw, L
通讯作者：
Liaw, L
DOI：
10.1517/14796694.1.1.37
发表时间：
2005-02-01
期刊：
Future oncology (London, England)
影响因子：
--
作者：
Chatterjee, Sabarni K;Zetter, Bruce R
通讯作者：
Zetter, Bruce R
影响因子：
4.8
作者：
Christensen, Brian;Kazanecki, Christian C.;Sorensen, Esben S.
通讯作者：
Sorensen, Esben S.