Preprint Communication Version 1 Preserved in Portico This version is not peer-reviewed

Development of Highly Sensitive Anti-Mouse CD39 Monoclonal Antibodies C39Mab-1 and C39Mab-2 for flow cytometry

Hiroyuki Suzuki
ORCID logo ,
Yuma Kudo
,
Mayuki Tawara
,
Nohara Goto
,
Kenichiro Ishikawa
,
Tsunenori Ouchida
ORCID logo ,
Tomohiro Tanaka
,
Teizo Asano
ORCID logo ,
Takuro Nakamura
,
Miyuki Yanaka
,
Mika K. Kaneko
,
Yukinari Kato
* ORCID logo
Version 1 : Received: 3 April 2023 / Approved: 4 April 2023 / Online: 4 April 2023 (02:29:25 CEST)

A peer-reviewed article of this Preprint also exists.

Okada, Y., Suzuki, H., Kaneko, M. K., & Kato, Y. (2024). Development of a Sensitive Anti-Mouse CD39 Monoclonal Antibody (C39Mab-1) for Flow Cytometry and Western Blot Analyses. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy. Okada, Y., Suzuki, H., Kaneko, M. K., & Kato, Y. (2024). Development of a Sensitive Anti-Mouse CD39 Monoclonal Antibody (C39Mab-1) for Flow Cytometry and Western Blot Analyses. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy.

Abstract

CD39 is involved in adenosine metabolism through conversion of extracellular ATP to adenosine. Because extracellular adenosine plays a critical role in the immune suppression of tumor microenvironment, the inhibition of CD39 activity by monoclonal antibodies (mAbs) is one of the important strategies for tumor therapy. In this study, we developed specific and sensitive mAbs for mouse CD39 (mCD39) using the Cell-Based Immunization and Screening (CBIS) method. The established anti-mCD39 mAbs, which were established by the CBIS method including C39Mab-1 (rat IgG2a, kappa) and C39Mab-2 (rat IgG2a, lambda), reacted with not only mCD39-overexpressed Chinese hamster ovary-K1 (CHO/mCD39) but also endogenously mCD39-expressed cell lines, such as L1210 (mouse lymphocytic leukemia) and J774-1 (mouse macrophage-like) cell lines through flow cytometry. Kinetic analyses using flow cytometry indicated that the dissociation constant (KD) of C39Mab-1 and C39Mab-2 for CHO/mCD39 was 7.3 × 10−9 M and 5.5 × 10−9 M, respectively. KD of C39Mab-1 and C39Mab-2 for L1210 was 3.3 × 10−9 M and 3.6 × 10−10 M, respectively. Furthermore, C39Mab-1 could detect the lysate of CHO/mCD39 by western blot analysis. These results indicate that C39Mab-1 and C39Mab-2 are useful for the detection of mCD39 in many functional studies.

Keywords

mouse CD39; monoclonal antibody; the Cell-Based Immunization and Screening; CBIS

Subject

Medicine and Pharmacology, Oncology and Oncogenics

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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