Establishment of Monoclonal Antibody PMab-202 Against Horse Podoplanin.
Abstract: Podoplanin (PDPN), a type I transmembrane glycoprotein, is expressed in several body tissues, including podocytes of renal glomerulus, type I alveolar cells of lung, and lymphatic endothelial cells. PDPN activates platelet aggregation by binding to C-type lectin-like receptor-2 (CLEC-2) presented on platelets. Monoclonal antibodies (mAbs) against human-, mouse-, rat-, rabbit-, dog-, bovine-, and cat-PDPN have already been established. However, anti-horse PDPN mAbs have not yet been developed. In this study, we immunized mice with synthetic horse PDPN peptides and developed anti-horse PDPN mAbs. One of the established mAbs, PMab-202 (IgG, kappa), was specifically able to detect horse PDPN in Chinese hamster ovary/horse PDPN (CHO/horPDPN) cells in flow cytometry experiments. PMab-202 was also able to detect endogenous horse PDPN expressed in and a horse kidney cell line, FHK-Tcl3.1, in flow cytometry and Western blot analyses. PMab-202 is expected to prove useful in investigating the function of horse PDPN.
Publication Date: 2018-10-27 PubMed ID: 30362932DOI: 10.1089/mab.2018.0030Google Scholar: Lookup
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- Journal Article
Summary
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This research outlines the development of a specific monoclonal antibody, PMab-202, designed to detect horse podoplanin, a protein important for platelet aggregation, which had not been created prior to this study.
Background on Podoplanin and Monoclonal Antibodies
- Podoplanin (PDPN) is a type 1 transmembrane glycoprotein expressed in various bodily tissues, such as renal glomerulus podocytes, lung’s type I alveolar cells, and lymphatic endothelial cells. It plays a crucial role in activating platelet aggregation by binding to the C-type lectin-like receptor-2 (CLEC-2) found on platelets.
- Monoclonal antibodies (mAbs) are a type of antibody made by identical immune cells, clones of a unique parent cell, designed to target a specific protein.
- Despite the development of monoclonal antibodies against PDPN in various species such as humans, mice, rats, rabbits, dogs, cows, and cats, none had been created specifically to target PDPN in horses before this study.
Purpose and Methodology of the Study
- The aim of this research was to develop an antibody that could specifically detect horse PDPN.
- The researchers accomplished this by immunizing mice with synthetic horse PDPN peptides, leading to the development of antibodies against horse PDPN.
Results of PMab-202 Development
- One of the resultant monoclonal antibodies, PMab-202, displayed the ability to specifically detect horse PDPN in particular cell types, as demonstrated by flow cytometry experiments.
- PMab-202 effectively detected endogenous horse PDPN expressed in and a horse kidney cell line, FHK-Tcl3.1, when evaluated through flow cytometry and Western blot analyses.
Future Implications of the Research
- PMab-202 is anticipated to be a valuable tool for studying the function of horse PDPN, widening the potential for more comprehensive research into the role of this protein in horses.
- This developed antibody also paves the way for further research looking at potential veterinary applications, particularly in the context of diseases or conditions where PDPN is implicated.
Cite This Article
APA
Furusawa Y, Yamada S, Itai S, Sano M, Nakamura T, Yanaka M, Handa S, Mizuno T, Maeda K, Fukui M, Harada H, Kaneko MK, Kato Y.
(2018).
Establishment of Monoclonal Antibody PMab-202 Against Horse Podoplanin.
Monoclon Antib Immunodiagn Immunother, 37(5), 233-237.
https://doi.org/10.1089/mab.2018.0030 Publication
Researcher Affiliations
- 1 Department of Antibody Drug Development, Tohoku University Graduate School of Medicine , Sendai, Miyagi, Japan .
- 2 New Industry Creation Hatchery Center, Tohoku University , Sendai, Miyagi, Japan .
- 3 ZENOAQ RESOURCE CO., LTD. , Koriyama, Fukushima, Japan .
- 1 Department of Antibody Drug Development, Tohoku University Graduate School of Medicine , Sendai, Miyagi, Japan .
- 1 Department of Antibody Drug Development, Tohoku University Graduate School of Medicine , Sendai, Miyagi, Japan .
- 4 Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University , Tokyo, Japan .
- 1 Department of Antibody Drug Development, Tohoku University Graduate School of Medicine , Sendai, Miyagi, Japan .
- 1 Department of Antibody Drug Development, Tohoku University Graduate School of Medicine , Sendai, Miyagi, Japan .
- 1 Department of Antibody Drug Development, Tohoku University Graduate School of Medicine , Sendai, Miyagi, Japan .
- 1 Department of Antibody Drug Development, Tohoku University Graduate School of Medicine , Sendai, Miyagi, Japan .
- 5 Laboratories of Molecular Diagnostics and Therapeutics, Joint Faculty of Veterinary Medicine, Yamaguchi University , Yamaguchi, Japan .
- 6 Laboratories of Veterinary Microbiology, Joint Faculty of Veterinary Medicine, Yamaguchi University , Yamaguchi, Japan .
- 3 ZENOAQ RESOURCE CO., LTD. , Koriyama, Fukushima, Japan .
- 4 Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University , Tokyo, Japan .
- 1 Department of Antibody Drug Development, Tohoku University Graduate School of Medicine , Sendai, Miyagi, Japan .
- 1 Department of Antibody Drug Development, Tohoku University Graduate School of Medicine , Sendai, Miyagi, Japan .
- 2 New Industry Creation Hatchery Center, Tohoku University , Sendai, Miyagi, Japan .
MeSH Terms
- Animals
- Antibodies, Monoclonal / biosynthesis
- Antibodies, Monoclonal / genetics
- Antibodies, Monoclonal / immunology
- Antibody Specificity / immunology
- CHO Cells
- Cricetinae
- Cricetulus
- Epitope Mapping
- Epitopes / immunology
- Flow Cytometry
- Horses / immunology
- Humans
- Lectins, C-Type / immunology
- Membrane Glycoproteins / genetics
- Membrane Glycoproteins / immunology
- Mice
- Platelet Aggregation / immunology
- Podocytes / immunology
Citations
This article has been cited 7 times.- Goto N, Suzuki H, Tanaka T, Ishikawa K, Ouchida T, Kaneko MK, Kato Y. EMab-300 Detects Mouse Epidermal Growth Factor Receptor-Expressing Cancer Cell Lines in Flow Cytometry.. Antibodies (Basel) 2023 Jun 21;12(3).
- Goto N, Suzuki H, Tanaka T, Asano T, Kaneko MK, Kato Y. Development of a Novel Anti-CD44 Monoclonal Antibody for Multiple Applications against Esophageal Squamous Cell Carcinomas.. Int J Mol Sci 2022 May 16;23(10).
- Suzuki H, Kaneko MK, Kato Y. Roles of Podoplanin in Malignant Progression of Tumor.. Cells 2022 Feb 7;11(3).
- Furusawa Y, Yamada S, Nakamura T, Sano M, Sayama Y, Itai S, Takei J, Harada H, Fukui M, Kaneko MK, Kato Y. PMab-235: A monoclonal antibody for immunohistochemical analysis against goat podoplanin.. Heliyon 2019 Jul;5(7):e02063.
- Kato Y, Furusawa Y, Yamada S, Itai S, Takei J, Sano M, Kaneko MK. Establishment of a monoclonal antibody PMab-225 against alpaca podoplanin for immunohistochemical analyses.. Biochem Biophys Rep 2019 Jul;18:100633.
- Furusawa Y, Yamada S, Itai S, Nakamura T, Takei J, Sano M, Harada H, Fukui M, Kaneko MK, Kato Y. Establishment of a monoclonal antibody PMab-233 for immunohistochemical analysis against Tasmanian devil podoplanin.. Biochem Biophys Rep 2019 Jul;18:100631.
- Furusawa Y, Yamada S, Itai S, Nakamura T, Yanaka M, Sano M, Harada H, Fukui M, Kaneko MK, Kato Y. PMab-219: A monoclonal antibody for the immunohistochemical analysis of horse podoplanin.. Biochem Biophys Rep 2019 Jul;18:100616.
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