Megakaryocyte potentiation factor cleaved from mesothelin precursor is a useful tumor marker in the serum of patients with mesothelioma
Megakaryocyte potentiation factor cleaved from mesothelin precursor is a useful tumor marker in the serum of patients with mesothelioma
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DOI：
10.1158/1078-0432.ccr-06-0472
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发表时间：
2006-07-15
影响因子：
11.5
通讯作者：
Pastan, Ira
中科院分区：
文献类型：
--
作者：
Onda, Masanori;Nagata, Satoshi;Pastan, Ira
目的：目的：建立抗巨核细胞增强因子（MPF）的单克隆抗体（mAb），并检测间皮瘤患者血液中MPF的表达。建立了几株产生MPF单克隆抗体的杂交瘤。一个双决定簇（夹心）ELISA构建使用单克隆抗体的两个不同的表位，并用于确定MPF是否存在于血清中的间皮瘤patients.Results：我们建立了7个抗MPF单克隆抗体的地形表位被分为三个不重叠的组。ELISA结果表明，所有mAb均能与重组MPF蛋白发生特异性反应。其中一种mAb在Western印迹上检测细胞裂解物中的MPF和含有MPF的间皮素前体蛋白。构建了使用针对两种不同表位的mAb的夹心ELISA，并用于测量各种表达间皮素的癌细胞系的培养基和人血清中MPF的存在。ELISA显示，与健康对照组相比，91%（56例中的51例）间皮瘤患者的MPF水平升高。此外，血清MPF下降到正常水平，在两个病人手术后，他们的腹膜间皮瘤。结论：使用新的单克隆抗体MPF，我们表明，MPF是由几个间皮瘤细胞系分泌，并经常升高间皮瘤患者的血液中。MPF的测量可能是有用的间皮瘤治疗后的反应。
Purpose: To establish monoclonal antibodies (mAb) against megakaryocyte potentiation factor (MPF) and detect MPF in the blood of patients with mesothelioma.Experimental Design: Mice were immunized with a purified recombinant human MPF-rabbit-Fc fusion protein and with MPF. Several hybridomas producing mAbs to MPF were established. A double-determinant (sandwich) ELISA was constructed using mAbs to two different epitopes and used to determine if MPF is present in the serum of patients with mesothelioma.Results: We established seven anti-MPF mAbs whose topographical epitopes were classified into three nonoverlapping groups. All the mAbs reacted with recombinant MPF protein by ELISA. One of the mAbs detected MPF and the mesothelin precursor protein containing MPF in cell lysates on Western blotting. A sandwich ELISA using mAbs to two different epitopes was constructed and used to measure the presence of MPF in the media of various mesothelin-expressing cancer cell lines and in human serum. The ELISA showed that MPF levels were elevated in 91% (51 of 56) of patients with mesothelioma compared with healthy controls. Furthermore, serum MPF fell to normal levels in two patients after surgery for their peritoneal mesothelioma.Conclusions: Using new mAbs to MPF, we showed that MPF is secreted by several mesothelioma cell lines and is frequently elevated in the blood of patients with mesothelioma. Measurement of MPF may be useful in following the response of mesothelioma to treatment.