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ID 118704
Title Alternative
PP-specific monoclonal antibodies
Author
Hara, Akemi Juntendo University
Nakagawa, Yuko Gunma University
Nakao, Keiko Saitama Medical University
Mizukami, Hiroki Hirosaki University
Maruyama, Nobuhiro Immuno-Biological Laboratories
Watada, Hirotaka Juntendo University
Fujitani, Yoshio Juntendo University|Gunma University
Keywords
Islet
Pancreatic polypeptide (PP)
Peptide YY
Neuropeptide Y
Monoclonal antibody
Content Type
Journal Article
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.
Journal Title
Endocrine Journal
ISSN
13484540
Publisher
The Japan Endocrine Society
Volume
66
Issue
5
Start Page
459
End Page
468
Published Date
2019
EDB ID
DOI (Published Version)
URL ( Publisher's Version )
FullText File
language
eng
TextVersion
Publisher
departments
Institute of Advanced Medical Sciences
University Hospital
Medical Sciences