Establishment of an antibody specific for cancer-associated haptoglobin: a possible implication of clinical investigation
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Kimihiro Nishino1,2,*, Sayaka Koda1,*, Naoya Kataoka1,*, Shinji Takamatsu1, Miyako Nakano3, Shun Ikeda1, Yuka Kamamatsu1, Koichi Morishita1, Kenta Moriwaki1, Hidetoshi Eguchi4, Eiko Yamamoto2,5, Fumitaka Kikkawa2, Yasuhiko Tomita6, Yoshihiro Kamada1 and Eiji Miyoshi1
1Department of Molecular Biochemistry and Clinical Investigation, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
2Department of Obstetrics and Gynecology, Nagoya University Graduate School of Medicine, Nagoya, Aichi, Japan
3Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashihiroshima, Hiroshima, Japan
4Department of Gastroenterological Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
5Department of Healthcare Administration, Nagoya University Graduate School of Medicine, Nagoya, Aichi, Japan
6Department of Pathology, International University of Health and Welfare, Narita, Chiba, Japan
*These authors contributed equally to this work
Eiji Miyoshi, email: firstname.lastname@example.org
Keywords: fucosylation; glycosylation; haptoglobin; glycan antibody; cancer biomarker
Received: May 24, 2017 Accepted: January 20, 2018 Published: January 29, 2018
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.
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