Hiroyuki Satofuka, Hiroyuki Suzuki, Mika K Kaneko, Yukinari Kato
{"title":"Development of Anti-Human Cadherin-26 Monoclonal Antibody, Ca<sub>26</sub>Mab-6, for Flow Cytometry.","authors":"Hiroyuki Satofuka, Hiroyuki Suzuki, Mika K Kaneko, Yukinari Kato","doi":"10.1177/21679436261428408","DOIUrl":"10.1177/21679436261428408","url":null,"abstract":"<p><p>Cadherin 26 (CDH26) is a recently identified member of the cadherin superfamily. Although <i>CDH26</i> gene expression has been reported in association with allergic inflammatory responses, the protein expression levels and the signaling pathways mediated through its interactions with other proteins remain poorly understood. This is primarily due to the lack of monoclonal antibodies (mAbs) that can recognize the intact, cell surface-expressed form of CDH26. In this study, we developed an antihuman CDH26 mAb, Ca<sub>26</sub>Mab-6 (IgM, kappa), using the Cell-Based Immunization and Screening (CBIS) method. Ca<sub>26</sub>Mab-6 demonstrated high sensitivity and specificity for CDH26 in flow cytometry and did not bind to Chinese hamster ovary (CHO)-K1 cells, which overexpress any of the other type I or type II cadherins. Ca<sub>26</sub>Mab-6 successfully detected endogenous CDH26 protein expression in HepG2, U-87 MG, MCF7, and 293FT cells. The apparent dissociation constant of Ca<sub>26</sub>Mab-6 was determined to be 9.8 ± 4.8 × 10<sup>-9</sup> M for CDH26-overexpressed CHO-K1 (CHO/CDH26) cells and 3.6 ± 1.0 × 10<sup>-7</sup> M for HepG2 cells. The detection of CDH26 expression in cancer cells may offer new insights into the potential relationship between inflammatory responses and malignant transformation. Therefore, Ca<sub>26</sub>Mab-6, developed using the CBIS method, is expected to facilitate functional studies of CDH26 and contribute to the development of CDH26-targeted antibody-based therapies.</p>","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"25-33"},"PeriodicalIF":0.0,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147391661","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Acknowledgment of Reviewers 2025.","authors":"","doi":"10.1177/21679436261431081","DOIUrl":"https://doi.org/10.1177/21679436261431081","url":null,"abstract":"","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"21679436261431081"},"PeriodicalIF":0.0,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147693503","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Honoka Taruta, Tomohiro Tanaka, Mika K Kaneko, Hiroyuki Suzuki, Yukinari Kato
{"title":"Cx<sub>2</sub>Mab-5: A Novel Anti-Mouse CXCR2 Monoclonal Antibody Developed by N-Terminus Peptide Immunization for Multiple Applications.","authors":"Honoka Taruta, Tomohiro Tanaka, Mika K Kaneko, Hiroyuki Suzuki, Yukinari Kato","doi":"10.1177/21679436251415261","DOIUrl":"10.1177/21679436251415261","url":null,"abstract":"<p><p>The CXC chemokine receptor 2 (CXCR2) is a member of the G-protein-coupled receptor superfamily and regulates a diverse range of immune responses and tumor progression. CXCR2 is expressed on immune cells, especially neutrophils, and is involved in various immune responses by interacting with its chemokine ligands. Therefore, the development of sensitive monoclonal antibodies (mAbs) for CXCR2 has been desired for treatment and diagnosis. This study established a novel sensitive anti-mouse CXCR2 (mCXCR2) mAb; Cx<sub>2</sub>Mab-5 (rat IgG<sub>2a</sub>, κ), using the mCXCR2 synthetic N-terminus peptide immunization method. In flow cytometry, Cx<sub>2</sub>Mab-5 recognized mCXCR2-overexpressed Chinese hamster ovary-K1 cells (CHO/mCXCR2) and WEHI-3B (murine myelomonocytic leukemia cell) cells, which express endogenous mCXCR2. Cx<sub>2</sub>Mab-5 did not cross-react with other mouse CC, CXC, CX3C, and XC chemokine receptors. Cx<sub>2</sub>Mab-5 showed a moderate binding affinity for both CHO/mCXCR2 and WEHI-3B. Furthermore, Cx<sub>2</sub>Mab-5 detected mCXCR2 in Western blot and immunohistochemistry in CHO/mCXCR2 cells, but a commercially available anti-mCXCR2 mAb (clone SA045E1) did not. Hence, Cx<sub>2</sub>Mab-5 can be a valuable tool for analyzing mCXCR2-positive cells in mouse tissues.</p>","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"11-20"},"PeriodicalIF":0.0,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146222087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Guanjie Li, Hiroyuki Suzuki, Mika K Kaneko, Yukinari Kato
{"title":"Establishment of Novel Anti-EphB3 Monoclonal Antibodies for Multiple Applications.","authors":"Guanjie Li, Hiroyuki Suzuki, Mika K Kaneko, Yukinari Kato","doi":"10.1177/21679436251408319","DOIUrl":"10.1177/21679436251408319","url":null,"abstract":"<p><p>Ephrin type-B receptor 3 (EphB3) binds to transmembrane ephrin-B ligands to regulate cell migration, adhesion, and proliferation. EphB3 exhibits a gradient expression pattern in the normal intestine, with the highest levels at the crypt base, and plays a crucial role in the maintenance of normal intestinal epithelium. Therefore, anti-EphB3 monoclonal antibodies (mAbs) are required for basic research and diagnosis. In this study, we developed novel antihuman EphB3, Eb<sub>3</sub>Mab-5 (IgG<sub>1</sub>, κ) and Eb<sub>3</sub>Mab-11 (IgG<sub>1</sub>, κ), using the Cell-Based Immunization and Screening (CBIS) method. Eb<sub>3</sub>Mab-5 and Eb<sub>3</sub>Mab-11 reacted with EphB3-overexpressed Chinese hamster ovary-K1 (CHO/EphB3) and endogenous EphB3-positive colorectal cancer LS174T in flow cytometry. The apparent binding affinity of Eb<sub>3</sub>Mab-5 for CHO/EphB3 and LS174T was 7.6 × 10<sup>-9</sup> M and 1.7 × 10<sup>-8</sup> M, respectively. Eb<sub>3</sub>Mab-11 could detect EphB3 in western blot analysis and immunohistochemistry. Eb<sub>3</sub>Mab-5 and Eb<sub>3</sub>Mab-11, established by the CBIS method, may contribute to the diagnosis and therapy of EphB3-positive tumors.</p>","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"1-10"},"PeriodicalIF":0.0,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145858974","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Generation of Rat Monoclonal Antibodies for Human and Mouse Androgen Receptor.","authors":"Momo Koike, Tomoka Kawabata, Norikazu Kiguchi, Yu Hatano, Kentaro Suzuki, Taro Tachibana, Chikako Yokoyama","doi":"10.1177/21679436251398987","DOIUrl":"10.1177/21679436251398987","url":null,"abstract":"<p><p>Androgen receptor (AR) is activated by binding to androgens, which leads to nuclear translocation, dimerization, and binding to androgen response elements (AREs) to regulate gene transcription. AR is important in masculinization during mammalian development and is a major driver of tumor growth in prostate cancer, for which AR pathway inhibitors are the standard treatment. However, the mechanisms by which AR participates in these processes remain unclear. In this study, we describe rat monoclonal antibodies (mAbs) that were generated against human and mouse AR. These mAbs recognize endogenous AR and were shown to be effective in the immunofluorescence staining of human cell lines and mouse tissue sections and in immunoprecipitation experiments. We expect these mAbs to be useful for functional analyses of AR.</p>","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"87-91"},"PeriodicalIF":0.0,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145558122","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Equitable Access to Antibody Libraries to Accelerate Discovery.","authors":"Andrei Moroz, Cory L Brooks","doi":"10.1177/21679436251404827","DOIUrl":"https://doi.org/10.1177/21679436251404827","url":null,"abstract":"","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":"44 6","pages":"85-86"},"PeriodicalIF":0.0,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145716540","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The Development of Monoclonal Antibodies Specific to the Soluble Form of CD155 in the Human Peripheral Blood.","authors":"Shota Kinoshita, Tomohei Matsuo, Soontae Gwon, Naoto Takeuchi, Akiko Iguchi-Manaka, Chigusa Nakahashi-Oda, Akira Shibuya, Kazuko Shibuya","doi":"10.1177/21679436251390201","DOIUrl":"https://doi.org/10.1177/21679436251390201","url":null,"abstract":"<p><p>CD155, also known as poliovirus receptor (PVR) or Necl-5, is an immunoreceptor with three immunoglobulin-like domains in the extracellular portion and is ubiquitously expressed on the cell surface of hematopoietic and nonhematopoietic cells. Human CD155 contains membrane-bound CD155 (mCD155) and soluble CD155 (sCD155) lacking the transmembrane region encoded by splicing variants of genes. The serum levels of sCD155 increased in patients with a variety of cancers. Furthermore, sCD155 suppresses tumor immunity through the blockade of DNAM-1 signaling. Therefore, sCD155 is potentially helpful for the diagnosis of cancer development and a novel therapeutic target in cancer treatment. However, monoclonal antibodies (mAbs) specific to sCD155, but not mCD155, have not yet been developed. Here, we generated 14 mAbs (named TX119 to TX122 and TX126 to TX135) that recognize the cytoplasmic region of CD155. These mAbs bind to sCD155, but not mCD155, when applied from the outside of the cell. Moreover, we established the assay system to quantify the sCD155 concentration in human serum. Thus, these mAbs can be utilized for the targeting and quantification of sCD155 in the human peripheral blood.</p>","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":"44 5","pages":"79-83"},"PeriodicalIF":0.0,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145294236","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Stunning Progress in <i>De Novo</i> Computationally Designed Antibodies.","authors":"Andrei Moroz, Cory L Brooks","doi":"10.1177/21679436251385401","DOIUrl":"10.1177/21679436251385401","url":null,"abstract":"","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"77-78"},"PeriodicalIF":0.0,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145202121","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Asad Saeed, Khurram Wajih Mehmood, Muhammad Irfan Ashiq, Sajjad Ahmad, Muhammad Sohail Anjum, Qaiser Hussain, Muhammad Amjad, Nauman Rauf
{"title":"Internal Validation of the Anti-Human Hemoglobin Antibody for Enhanced Human Blood Detection at the PFSA DNA and Serology Laboratory, Pakistan.","authors":"Asad Saeed, Khurram Wajih Mehmood, Muhammad Irfan Ashiq, Sajjad Ahmad, Muhammad Sohail Anjum, Qaiser Hussain, Muhammad Amjad, Nauman Rauf","doi":"10.1089/mab.2025.0002","DOIUrl":"10.1089/mab.2025.0002","url":null,"abstract":"<p><p>Biological fluids collected from crime scenes play a crucial role in solving serious crimes such as murder, rape, burglary, and theft. Identifying human blood using various methods is crucial for linking disparate pieces of evidence and solving crimes. In this study, the Anti-Human Hemoglobin antibody (Ah-HB) from Sigma-Aldrich® USA was internally validated for human blood identification using the Ouchterlony Double Immunodiffusion (ODD) technique at the DNA and Serology Department of the Punjab Forensic Science Agency in Lahore, Pakistan. Additionally, a comparative analysis was conducted with Seratec® HemDirect (S_HD) strips to evaluate the economic feasibility of both methods. The internal validation included assessments of sensitivity, specificity, and an analysis of real-case work samples to determine the viability of the antibody as a confirmatory test for human blood. Although Ah-HB had lower sensitivity in detecting human blood at higher dilutions (1:10,000), it offered a more cost-effective solution per sample compared with S_HD, which demonstrated higher sensitivity (1:2,000,000) but at a significantly higher cost per sample. Specificity tests revealed no cross-reactivity with nonhuman blood for both Ah-HB and S_HD. This study emphasizes the importance of selecting suitable antibodies for forensic analysis by evaluating sensitivity, specificity, and cost-effectiveness. Ah-HB emerges as a valuable tool for forensic laboratories, providing reliable results at a lower cost compared with S_HD, thereby enhancing the efficiency and effectiveness of criminal investigations.</p>","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"71-76"},"PeriodicalIF":0.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144555713","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}