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Establishment of an antibody specific for cancer-associated haptoglobin: a possible implication of clinical investigation

2018; Impact Journals LLC; Volume: 9; Issue: 16 Linguagem: Inglês

10.18632/oncotarget.24332

1949-2553

Kimihiro Nishino, Sayaka Koda, Naoya Kataoka, Shinji Takamatsu, Miyako Nakano, Shun Ikeda, Yuka Kamamatsu, Koichi Morishita, Kenta Moriwaki, Hidetoshi Eguchi, Eiko Yamamoto, Fumitaka Kikkawa, Yasuhiko Tomita, Yoshihiro Kamada, Eiji Miyoshi,

Erythrocyte Function and Pathophysiology

// Kimihiro Nishino 1, 2, * , Sayaka Koda 1, * , Naoya Kataoka 1, * , Shinji Takamatsu 1 , Miyako Nakano 3 , Shun Ikeda 1 , Yuka Kamamatsu 1 , Koichi Morishita 1 , Kenta Moriwaki 1 , Hidetoshi Eguchi 4 , Eiko Yamamoto 2, 5 , Fumitaka Kikkawa 2 , Yasuhiko Tomita 6 , Yoshihiro Kamada 1 and Eiji Miyoshi 1 1 Department of Molecular Biochemistry and Clinical Investigation, Osaka University Graduate School of Medicine, Suita, Osaka, Japan 2 Department of Obstetrics and Gynecology, Nagoya University Graduate School of Medicine, Nagoya, Aichi, Japan 3 Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashihiroshima, Hiroshima, Japan 4 Department of Gastroenterological Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan 5 Department of Healthcare Administration, Nagoya University Graduate School of Medicine, Nagoya, Aichi, Japan 6 Department of Pathology, International University of Health and Welfare, Narita, Chiba, Japan * These authors contributed equally to this work Correspondence to: Eiji Miyoshi, email: emiyoshi@sahs.med.osaka-u.ac.jp Keywords: fucosylation; glycosylation; haptoglobin; glycan antibody; cancer biomarker Received: May 24, 2017 Accepted: January 20, 2018 Published: January 29, 2018 ABSTRACT 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.