Version 1
: Received: 18 July 2024 / Approved: 19 July 2024 / Online: 19 July 2024 (06:39:45 CEST)
How to cite:
Hirose, M.; Suzuki, H.; Ubukata, R.; Tanaka, T.; Kaneko, M. K.; Kato, Y. Development of Specific Anti-mouse Atypical Chemokine Receptor 4 Monoclonal Antibodies. Preprints2024, 2024071576. https://doi.org/10.20944/preprints202407.1576.v1
Hirose, M.; Suzuki, H.; Ubukata, R.; Tanaka, T.; Kaneko, M. K.; Kato, Y. Development of Specific Anti-mouse Atypical Chemokine Receptor 4 Monoclonal Antibodies. Preprints 2024, 2024071576. https://doi.org/10.20944/preprints202407.1576.v1
Hirose, M.; Suzuki, H.; Ubukata, R.; Tanaka, T.; Kaneko, M. K.; Kato, Y. Development of Specific Anti-mouse Atypical Chemokine Receptor 4 Monoclonal Antibodies. Preprints2024, 2024071576. https://doi.org/10.20944/preprints202407.1576.v1
APA Style
Hirose, M., Suzuki, H., Ubukata, R., Tanaka, T., Kaneko, M. K., & Kato, Y. (2024). Development of Specific Anti-mouse Atypical Chemokine Receptor 4 Monoclonal Antibodies. Preprints. https://doi.org/10.20944/preprints202407.1576.v1
Chicago/Turabian Style
Hirose, M., Mika K Kaneko and Yukinari Kato. 2024 "Development of Specific Anti-mouse Atypical Chemokine Receptor 4 Monoclonal Antibodies" Preprints. https://doi.org/10.20944/preprints202407.1576.v1
Abstract
Leukocyte migration is an essential function of innate and adaptive immune responses. Chemokines and their receptors control the migration system. The abundance of chemokines is controlled by atypical chemokine receptors (ACKRs), chemokine receptor-like molecules that do not couple to the G protein signaling pathways. Among them, ACKR4 regulates dendritic cell migration by controlling the ligands and is involved in tumor development in mouse models. Because no anti-mACKR4 mAb for flow cytometry has been reported, this study aimed to develop a novel monoclonal antibody (mAb) for mouse ACKR4 (mACKR4). Among the established anti-mACKR4 mAbs, A4Mab-1 (rat IgG2b, kappa), A4Mab-2 (rat IgG2b, kappa), and A4Mab-3 (rat IgG2b, kappa) recognized mACKR4-overexpressed Chinese hamster ovary-K1 (CHO/mACKR4) by flow cytometry. The dissociation constant (KD) values of A4Mab-1, A4Mab-2, and A4Mab-3 for CHO/mACKR4 were determined as 3.5 × 10−9 M, 6.0 × 10−9 M, and 1.6 × 10−9 M, respectively. Furthermore, A4Mab-1 and A4Mab-2 could detect mACKR4 by western blotting. These results indicated that A4Mab-1, A4Mab-2, and A4Mab-3 help to detect mACKR4 by flow cytometry and western blotting and obtain the proof of concept in preclinical models.
Keywords
mouse ACKR4; monoclonal antibody; peptide immunization; flow cytometry; western blotting
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.
