Development of monoclonal mouse antibodies that specifically recognize pancreatic polypeptide

Access this Article

Author(s)

    • Hara Akemi
    • Department of Metabolism & Endocrinology, Juntendo University Graduate School of Medicine, Tokyo 113-8421, Japan|Center for Therapeutic Innovation in Diabetes, Juntendo University Graduate School of Medicine, Tokyo 113-8421, Japan
    • Watada Hirotaka
    • Department of Metabolism & Endocrinology, Juntendo University Graduate School of Medicine, Tokyo 113-8421, Japan|Center for Therapeutic Innovation in Diabetes, Juntendo University Graduate School of Medicine, Tokyo 113-8421, Japan|Center for Identification of Diabetic Therapeutic Targets, Juntendo University Graduate School of Medicine, Tokyo 113-8421, Japan|Sportology Center, Juntendo University Graduate School of Medicine, Tokyo 113-8421, Japan
    • Fujitani Yoshio
    • Department of Metabolism & Endocrinology, Juntendo University Graduate School of Medicine, Tokyo 113-8421, Japan|Center for Therapeutic Innovation in Diabetes, Juntendo University Graduate School of Medicine, Tokyo 113-8421, Japan|Laboratory of Developmental Biology & Metabolism, Institute for Molecular and Cellular Regulation, Gunma University, Gunma 371-8512, Japan
    • Nakagawa Yuko
    • Laboratory of Developmental Biology & Metabolism, Institute for Molecular and Cellular Regulation, Gunma University, Gunma 371-8512, Japan
    • Nakao Keiko
    • Department of Physiology, Faculty of Medicine, Saitama Medical University, Saitama 350-0495, Japan
    • Tamaki Motoyuki
    • Diabetes Therapeutics and Research Center, Institute of Advanced Medical Sciences, Tokushima University, Tokushima 770-8503, Japan
    • Ikemoto Tetsuya
    • Department of Digestive and Transplant Surgery, Tokushima University, Tokushima 770-8503, Japan
    • Shimada Mitsuo
    • Department of Digestive and Transplant Surgery, Tokushima University, Tokushima 770-8503, Japan
    • Matsuhisa Munehide
    • Diabetes Therapeutics and Research Center, Institute of Advanced Medical Sciences, Tokushima University, Tokushima 770-8503, Japan
    • Mizukami Hiroki
    • Department of Pathology and Molecular Medicine, Hirosaki University Graduate School of Medicine, Aomori 036-8562, Japan

Abstract

<p>Pancreatic polypeptide (PP) is a 36-amino acid peptide encoded by the <i>Ppy</i> 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 <i>Ppy</i> knockout (<i>Ppy</i>-KO) mice in which the <i>Ppy</i>-coding region was replaced by Cre recombinase. The <i>Ppy</i>-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 <i>Ppy</i>-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.</p>

Journal

  • Endocrine Journal

    Endocrine Journal 66(5), 459-468, 2019

    The Japan Endocrine Society

Codes

Page Top