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Establishment and characterization of a fucosylated α-fetoprotein-specific monoclonal antibody: a potential application for clinical research.
Scientific Reports ( IF 3.8 ) Pub Date : 2019-08-26 , DOI: 10.1038/s41598-019-48821-x Yuriko Egashira 1 , Masatoshi Suganuma 2 , Yukiko Kataoka 1 , Yukiko Higa 1 , Nobuyuki Ide 3 , Koichi Morishita 4 , Yoshihiro Kamada 4 , Jianguo Gu 5 , Koji Fukagawa 1 , Eiji Miyoshi 4
Scientific Reports ( IF 3.8 ) Pub Date : 2019-08-26 , DOI: 10.1038/s41598-019-48821-x Yuriko Egashira 1 , Masatoshi Suganuma 2 , Yukiko Kataoka 1 , Yukiko Higa 1 , Nobuyuki Ide 3 , Koichi Morishita 4 , Yoshihiro Kamada 4 , Jianguo Gu 5 , Koji Fukagawa 1 , Eiji Miyoshi 4
Affiliation
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The Lens culinaris agglutinin (LCA)-reactive fraction of α-fetoprotein (AFP-L3) is a well-known cancer biomarker for hepatocellular carcinoma (HCC) with very high specificity. Because LCA recognizes only bi-antennary N-glycans with a core fucose, some of fucosylated AFP in HCC patients may not be detected. Then glycan antibodies, which recognize both specific glycan and protein, are desired for glycobiology. Here, we successfully established a novel glycan antibody for fucosylated AFP and demonstrated its potential clinical application. After immunization with a fucosylated AFP peptide, positive screening was performed for fucosylated AFP peptides using solid-phase enzyme-linked immunosorbent assay (ELISA). The newly developed antibody was designated: fucosylated AFP-specific mAb (FasMab). Western blot analysis showed that FasMab reacted with AFP produced by HepG2 cells, but not with AFP produced by α-1,6-fucosyltransferase deficient HepG2 cells. The specific binding of FasMab to fucosylated AFP was confirmed with ELISA as well as western blot analysis. A preliminary high sensitivity chemiluminescence enzyme immunoassay kit showed increased levels of fucosylated AFP in the sera of patients with HCC, but not in the sera of normal patients, or patients with chronic liver diseases. Thus, the novel glycan antibody, FasMab, is a promising tool to study fucosylated AFP with clinical and basic research applications.
中文翻译：
岩藻糖基化的甲胎蛋白特异性单克隆抗体的建立和表征：临床研究的潜在应用。
扁豆凝集素（LCA）的α-甲胎蛋白（AFP-L3）反应级分是肝细胞癌（HCC）的知名癌症生物标志物，具有很高的特异性。由于LCA仅识别具有核心岩藻糖的双天线N聚糖，因此可能未检测到HCC患者中一些岩藻糖基化的AFP。因此，糖原生物学需要同时识别特异性聚糖和蛋白质的聚糖抗体。在这里，我们成功地建立了岩藻糖基化AFP的新型聚糖抗体，并证明了其潜在的临床应用。用岩藻糖基化的AFP肽免疫后，使用固相酶联免疫吸附测定（ELISA）对岩藻糖基化的AFP肽进行阳性筛选。新开发的抗体命名为：岩藻糖基化的AFP特异性mAb（FasMab）。蛋白质印迹分析表明，FasMab与HepG2细胞产生的AFP反应，但不与α-1,6-岩藻糖基转移酶缺陷的HepG2细胞产生的AFP反应。ELISA和蛋白质印迹分析证实了FasMab与岩藻糖基化AFP的特异性结合。初步的高灵敏度化学发光酶免疫分析试剂盒显示，肝癌患者血清中岩藻糖基化的AFP水平升高，而正常患者或慢性肝病患者的血清中岩藻糖基化AFP水平却未升高。因此，新型的聚糖抗体FasMab是具有临床和基础研究应用的研究岩藻糖基化AFP的有前途的工具。初步的高灵敏度化学发光酶免疫分析试剂盒显示，肝癌患者血清中岩藻糖基化的AFP水平升高，而正常患者或慢性肝病患者的血清中岩藻糖基化AFP水平却未升高。因此，新型的聚糖抗体FasMab是具有临床和基础研究应用的研究岩藻糖基化AFP的有前途的工具。初步的高灵敏度化学发光酶免疫分析试剂盒显示，肝癌患者血清中岩藻糖基化的AFP水平升高，而正常患者或慢性肝病患者的血清中岩藻糖基化AFP水平却未升高。因此，新型的聚糖抗体FasMab是具有临床和基础研究应用的研究岩藻糖基化AFP的有前途的工具。
更新日期：2019-08-26
中文翻译：
岩藻糖基化的甲胎蛋白特异性单克隆抗体的建立和表征：临床研究的潜在应用。
扁豆凝集素（LCA）的α-甲胎蛋白（AFP-L3）反应级分是肝细胞癌（HCC）的知名癌症生物标志物，具有很高的特异性。由于LCA仅识别具有核心岩藻糖的双天线N聚糖，因此可能未检测到HCC患者中一些岩藻糖基化的AFP。因此，糖原生物学需要同时识别特异性聚糖和蛋白质的聚糖抗体。在这里，我们成功地建立了岩藻糖基化AFP的新型聚糖抗体，并证明了其潜在的临床应用。用岩藻糖基化的AFP肽免疫后，使用固相酶联免疫吸附测定（ELISA）对岩藻糖基化的AFP肽进行阳性筛选。新开发的抗体命名为：岩藻糖基化的AFP特异性mAb（FasMab）。蛋白质印迹分析表明，FasMab与HepG2细胞产生的AFP反应，但不与α-1,6-岩藻糖基转移酶缺陷的HepG2细胞产生的AFP反应。ELISA和蛋白质印迹分析证实了FasMab与岩藻糖基化AFP的特异性结合。初步的高灵敏度化学发光酶免疫分析试剂盒显示，肝癌患者血清中岩藻糖基化的AFP水平升高，而正常患者或慢性肝病患者的血清中岩藻糖基化AFP水平却未升高。因此，新型的聚糖抗体FasMab是具有临床和基础研究应用的研究岩藻糖基化AFP的有前途的工具。初步的高灵敏度化学发光酶免疫分析试剂盒显示，肝癌患者血清中岩藻糖基化的AFP水平升高，而正常患者或慢性肝病患者的血清中岩藻糖基化AFP水平却未升高。因此，新型的聚糖抗体FasMab是具有临床和基础研究应用的研究岩藻糖基化AFP的有前途的工具。初步的高灵敏度化学发光酶免疫分析试剂盒显示，肝癌患者血清中岩藻糖基化的AFP水平升高，而正常患者或慢性肝病患者的血清中岩藻糖基化AFP水平却未升高。因此，新型的聚糖抗体FasMab是具有临床和基础研究应用的研究岩藻糖基化AFP的有前途的工具。
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