ID | 118704 |
Title Alternative | PP-specific monoclonal antibodies
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Author |
Hara, Akemi
Juntendo University
Nakagawa, Yuko
Gunma University
Nakao, Keiko
Saitama Medical University
Ikemoto, Tetsuya
Tokushima University
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Shimada, Mitsuo
Tokushima University
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Matsuhisa, Munehide
Tokushima University
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Mizukami, Hiroki
Hirosaki University
Maruyama, Nobuhiro
Immuno-Biological Laboratories
Watada, Hirotaka
Juntendo University
Fujitani, Yoshio
Juntendo University|Gunma University
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Keywords | Islet
Pancreatic polypeptide (PP)
Peptide YY
Neuropeptide Y
Monoclonal antibody
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Content Type |
Journal Article
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Description | Pancreatic polypeptide (PP) is a 36-amino acid peptide encoded by the Ppy gene, which is produced by a small population of cells located in the periphery of the islets of Langerhans. Owing to the high amino acid sequence similarity among neuropeptide Y family members, antibodies against PP that are currently available are not convincingly specific to PP. Here we report the development of mouse monoclonal antibodies that specifically bind to PP. We generated Ppy knockout (Ppy-KO) mice in which the Ppy-coding region was replaced by Cre recombinase. The Ppy-KO mice were immunized with mouse PP peptide, and stable hybridoma cell lines producing anti-PP antibodies were isolated. Firstly, positive clones were selected in an enzyme-linked immunosorbent assay for reactivity with PP coupled to bovine serum albumin. During the screening, hybridoma clones producing antibodies that cross-react to the peptide YY (PYY) were excluded. In the second screening, hybridoma clones in which their culture media produce no signal in Ppy-KO islets but detect specific cells in the peripheral region of wild-type islets, were selected. Further studies demonstrated that the selected monoclonal antibody (23-2D3) specifically recognizes PP-producing cells, not only in mouse, but also in human and rat islets. The monoclonal antibodies with high binding specificity for PP developed in this study will be fundamental for future studies towards elucidating the expression profiles and the physiological roles of PP.
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Journal Title |
Endocrine Journal
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ISSN | 13484540
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Publisher | The Japan Endocrine Society
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Volume | 66
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Issue | 5
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Start Page | 459
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End Page | 468
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Published Date | 2019
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EDB ID | |
DOI (Published Version) | |
URL ( Publisher's Version ) | |
FullText File | |
language |
eng
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TextVersion |
Publisher
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departments |
Institute of Advanced Medical Sciences
University Hospital
Medical Sciences
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