Establishment of an antibody specific for cancer-associated haptoglobin: a possible implication of clinical investigation.
Establishment of an antibody specific for cancer-associated haptoglobin: a possible implication of clinical investigation.
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DOI：
10.18632/oncotarget.24332
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发表时间：
2018-02-27
期刊：
影响因子：
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通讯作者：
Miyoshi E
中科院分区：
文献类型：
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作者：
Nishino K;Koda S;Kataoka N;Takamatsu S;Nakano M;Ikeda S;Kamamatsu Y;Morishita K;Moriwaki K;Eguchi H;Yamamoto E;Kikkawa F;Tomita Y;Kamada Y;Miyoshi E
我们先前发现胰腺癌患者血清岩藻糖化结合珠蛋白(Fuc-HPT)水平显著升高。为了阐明这种增加的机制并开发一种简单的检测方法，我们着手制备一种针对Fuc-HPT的单抗。经酶联免疫吸附试验(EL ISA)多次筛选，10-7G单抗被鉴定为对细胞系产生的Fuc-HPT和胰腺癌患者的HPT具有高度特异性。经10-7G单抗亲和层析、凝集素印迹和质谱分析，发现10-7G单抗主要识别Fuc-HPT和也是岩藻糖化的前结合珠蛋白(ProHpt)。在免疫组织化学分析中，转移癌细胞周围的肝细胞被10-7G单抗染色，而原癌细胞本身和正常肝细胞均未见阳性染色，提示转移癌细胞促进邻近肝细胞产生Fuc-HPT。用自制的10-7G单抗建立了检测胰腺癌和结直肠癌患者血清Fuc-HPT水平的新方法。有趣的是，在结直肠癌IV期观察到Fuc-HPT水平显著增加。这些结果表明，所研制的10-7G单抗是一种很有前途的识别Fuc-HPT的抗体，有望成为检测肿瘤肝转移的有用工具。
We previously found that the serum level of fucosylated haptoglobin (Fuc-Hpt) was significantly increased in pancreatic cancer patients. To delineate the mechanism underlying this increase and develop a simple detection method, we set out to generate a monoclonal antibody (mAb) specific for Fuc-Hpt. After multiple screenings by enzyme-linked immunosorbent assay (ELISA), a 10-7G mAb was identified as being highly specific for Fuc-Hpt generated in a cell line as well as for Hpt derived from a pancreatic cancer patient. As a result from affinity chromatography with 10-7G mAb, followed by lectin blot and mass spectrometry analyses, it was found that 10-7G mAb predominantly recognized both Fuc-Hpt and prohaptoglobin (proHpt), which was also fucosylated. In immunohistochemical analyses, hepatocytes surrounding metastasized cancer cells were stained by the 10-7G mAb, but neither the original cancer cells themselves nor normal hepatocytes exhibited positive staining, suggesting that metastasized cancer cells promote Fuc-Hpt production in adjacent hepatocytes. Serum level of Fuc-Hpt determined with newly developed ELISA system using the 10-7G mAb, was increased in patients of pancreatic and colorectal cancer. Interestingly, dramatic increases in Fuc-Hpt levels were observed at the stage IV of colorectal cancer. These results indicate that the 10-7G mAb developed is a promising antibody which recognizes Fuc-Hpt and could be a useful diagnostic tool for detecting liver metastasis of cancer.