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Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

Mika K Kaneko, Shinji Yamada, Shunsuke Itai, Yao-Wen Chang, Takuro Nakamura, Miyuki Yanaka, Hiroyuki Harada, Hiroyoshi Suzuki, Yukinari Kato
Telomerase reverse transcriptase (TERT) and mutations of the TERT promoter are significant in the pathogenesis of 1p/19q-codeleted oligodendrogliomas and isocitrate dehydrogenase gene wild-type glioblastomas, as well as melanomas and squamous cell carcinomas. We previously developed an antihuman TERT monoclonal antibody (mAb), TMab-6, which is applicable in immunohistochemistry for human tissues. However, the binding epitope of TMab-6 against TERT is yet to be elucidated. In this study, enzyme-linked immunosorbent assay and immunohistochemistry were utilized for investigating the epitope of TMab-6...
May 3, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Yuho Nakamura, Keisuke Naito, Yumi Yamashita-Kanemaru, Daisuke Komori, Rei Hirochika, Akira Shibuya, Kazuko Shibuya
T cell immunoglobulin and ITIM domains (TIGIT) is an inhibitory immunoreceptor expressed on NK cells, effector and memory T cells, and regulatory T cells (Tregs). The ligands for TIGIT are CD155 (PVR) and CD112 (PVRL2, nectin-2), which are broadly expressed on hematopoietic cells and nonhematopoietic cells. TIGIT negatively regulates antitumor responses, but promotes autoimmune reaction. Although neutralizing anti-human TIGIT mAbs are under clinical trials for cancers, how the blockade of TIGIT interaction with the ligands shows tumor immunity still remains unclear...
April 12, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Balraj Palani
Malaria is a life-threatening infectious disease and continues to be a major public health crisis in many parts of the tropical world. Plasmodium falciparum is responsible for the majority of mortality and morbidity associated with malaria. During the intraerythrocytic cycle, P. falciparum releases three proteins with high histidine content as follows: histidine-rich protein 1 (HRP1), histidine-rich protein 2 (HRP2), and histidine-rich protein 3 (HRP3). Currently, most of the diagnostic tests of P. falciparum infection target HRP2, and a number of monoclonal antibodies (mAbs) against HRP2 have been developed for use in HRP2 detection and quantification...
April 10, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Qi Wang, Shuai-Yong Wang, Peng Zhang, Xiao-Min Liu, Ling-Xue Yu, Tong-Ling Shan, Wu Tong, Yan-Jun Zhou, Guo-Xin Li, Hao Zheng, Fei Gao, Yi-Feng Jiang, Ning Kong, Li-Wei Li, Guang-Zhi Tong, Hai Yu
The purified whole-virus proteins derived from A/swine/Shanghai/1/2014 (H1N1) (SH1) were chosen to immunize BALB/c mice to prepare the monoclonal antibody (MAb) against hemagglutinin (HA) protein of an European avian-like (EA) H1N1 swine influenza virus (SIV). After cloning three times by limiting dilution, one strain of hybridoma cells named 3C7 secreting anti-HA protein MAb was obtained by hybridoma technique. The results of indirect immunofluorescence assay and western blot analyses showed that the MAb 3C7 specifically reacted with the HA protein of EA H1N1 SIV...
April 9, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Mika K Kaneko, Shunsuke Itai, Shinji Yamada, Yukinari Kato
Esophageal cancer is one of the highly malignant cancers. It comprises two of the most common histological tumor types: squamous cell carcinoma (SCC) and adenocarcinoma. SCC accounts for about 90% of esophageal cancers. Despite developments in treatment strategies, the prognosis and survival rate remain poor. Podocalyxin (PODXL) is a highly glycosylated type-I transmembrane protein. It is expressed in normal tissues such as kidney, heart, breast, and pancreas. Upregulation of PODXL correlates with tumor progression, invasion, and metastasis...
April 9, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Shinji Yamada, Shunsuke Itai, Takuro Nakamura, Miyuki Yanaka, Yao-Wen Chang, Hiroyoshi Suzuki, Mika K Kaneko, Yukinari Kato
Programmed cell death ligand-1 (PD-L1) is a type I transmembrane glycoprotein expressed on antigen-presenting cells. It is also expressed in several tumor cells such as melanoma and lung cancer cells. A strong correlation has been reported between human PD-L1 (hPD-L1) expression in tumor cells and negative prognosis in cancer patients. Here, a novel anti-hPD-L1 monoclonal antibody (mAb) L1 Mab-13 (IgG1 , kappa) was produced using a cell-based immunization and screening (CBIS) method. We investigated hPD-L1 expression in lung cancer using flow cytometry, Western blot, and immunohistochemical analyses...
April 2, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Shinji Yamada, Mika K Kaneko, Shunsuke Itai, Yao-Wen Chang, Takuro Nakamura, Miyuki Yanaka, Satoshi Ogasawara, Takeshi Murata, Hiroaki Uchida, Hideaki Tahara, Hiroyuki Harada, Yukinari Kato
Podoplanin (PDPN), a type I transmembrane sialoglycoprotein, is expressed on normal renal podocytes, pulmonary type I alveolar cells, and lymphatic endothelial cells. Increased expression of PDPN in cancers is associated with poor prognosis and hematogenous metastasis through interactions with C-type lectin-like receptor 2 (CLEC-2) on platelets. We previously reported a novel PMab-48 antibody, which is an anti-dog PDPN (dPDPN) monoclonal antibody (mAb) recognizing PDPN expressed in lymphatic endothelial cells...
April 2, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Shinji Yamada, Shunsuke Itai, Mika K Kaneko, Yukinari Kato
Lung cancer is one of the leading causes of cancer-related deaths in the world. Regardless of the advances in lung cancer treatments, the prognosis is still poor. Podocalyxin (PODXL) is a highly glycosylated type I transmembrane protein that is expressed in normal tissues, including the heart, pancreas, and breast. It is also found and used as a diagnostic marker in many cancers, such as renal, brain, breast, oral, and lung cancers. We previously developed specific and sensitive anti-PODXL monoclonal antibodies, PcMab-47 (mouse IgG1 , kappa) and its mouse IgG2a -type (47-mG2a ), both of which were suitable for immunohistochemical analyses of oral cancers...
March 14, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Rodrigo de Almeida, Cecília Naomi Nakamura, Marina de Lima Fontes, Julia Paiola da Silva, Matheus Bertanha, Elenice Deffune, Ana Marisa Fusco-Almeida, Andrei Moroz
We report an immunization technique that can update the production of monoclonal antibodies (mAbs): the multiple tolerization subtractive immunization (MTSI). A total of 10 BALB/C mice were used. Animals in group 1 received one inoculation of RWPE-1 cells (nontumoral), followed by cyclophosphamide, and then received serial inoculations of nonirradiated PC3 cells (tumoral). Animals in group 2 received our MTSI protocol, as follows: one inoculation of RWPE-1 cells, followed by cyclophosphamide (Cy). This whole tolerization step was repeated three other times, with 14-day intervals between the last Cy exposure and the next RWPE-1 cell inoculation...
April 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Liwei Li, Fei Gao, Yifeng Jiang, Wu Tong, Hao Zheng, Tongling Shan, Ning Kong, Hai Yu, Deqiang Yang, Kuan Zhao, Yujiao Zhang, Guangzhi Tong
Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important viral pathogens that has caused tremendous economic losses to the swine industry worldwide. Although extensive research has been focused on PRRSV, little is known about the structure and biological functions of individual nonstructural viral proteins, especially the nonstructural protein 12 (Nsp12). In this study, we generated and identified the monoclonal antibody (mAb) against PRRSV Nsp12. Six strains of hybridoma cells named 2B10, 2B12, 5E1, 5G6, 5E7, and 8B2 secreting anti-Nsp12 mAbs were obtained by the hybridoma technique...
April 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Barbara Manfredi, Jessica Morales-Ortíz, Lymarie M Díaz-Díaz, Liz Hernandez-Matias, Delmaliz Barreto-Vázquez, Javier Menéndez-Pérez, J Alejandro Rodríguez-Cordero, Juan C Villalobos-Santos, Edgardo Santiago-Rivera, Adriana Rivera-Dompenciel, Eunice L Lozada-Delgado, Madhavi Kuchibhotla, Kelvin Carrasquillo-Carrión, Abiel Roche-Lima, A Valance Washington
Platelets play a vital role in hemostasis and inflammation. The membrane receptor TREM-like transcript-1 (TLT-1) is involved in platelet aggregation, bleeding, and inflammation, and it is localized in the α-granules of platelets. Upon platelet activation, TLT-1 is released from α-granules both in its transmembrane form and as a soluble fragment (sTLT-1). Higher levels of sTLT-1 have been detected in the plasma of patients with acute inflammation or sepsis, suggesting an important role for TLT-1 during inflammation...
April 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Yao-Wen Chang, Mika K Kaneko, Shinji Yamada, Yukinari Kato
The mucin-type membrane glycoprotein podoplanin (PDPN) is frequently overexpressed in numerous malignant cancers, including squamous cell carcinoma, germinal neoplasia, mesothelioma, lung cancer, oral cancer, and brain tumor. PDPN expression is strongly associated with cancer progression and poor prognosis. Furthermore, PDPN binds to C-type lectin-like receptor 2 (CLEC-2) on platelets, followed by PDPN-mediated platelet aggregation to facilitate tumor metastasis. We have previously reported a novel anti-cat PDPN (cPDPN) monoclonal antibody (mAb), PMab-52, which specifically detects cPDPN using flow cytometry analysis and successfully identifies cPDPN in feline squamous cell carcinomas...
February 2, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Osamu Kusano-Arai, Hiroko Iwanari, Shota Kudo, Chika Kikuchi, Anna Yui, Hiroki Akiba, Keisuke Matsusaka, Atsushi Kaneda, Masashi Fukayama, Kouhei Tsumoto, Takao Hamakubo
Cadherin-17 (CDH17) is highly expressed in gastric cancer and is thus considered to be a good target for antibody therapy. CDH17 is classified as a nonclassical cadherin, in that it is composed of seven extracellular cadherin domains. We generated anti-CDH17 monoclonal antibodies (mAbs) which recognize the extracellular domain of CDH17. Competitive assay using AGS, a gastric cancer cell line, cells revealed that five selected anti-CDH17 mAbs recognize different epitopes on CDH17. As AGS cells were shown to exhibit broad expression pattern of CDH17 by flow cytometry, we separated three clones with a low (10,000/cell), medium (50,000/cell), and high (200,000/cell) expression level, designating them as AGSlow , AGSmed , and AGShigh , respectively...
February 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
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February 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Thomas P Conway, Karla J Daniels, Yang-Nim Park, David R Soll
Firefly luciferase (FLuc) is commonly used as a reporter gene PpyLuc1 in bioanalytical assays. We have produced five mouse-derived monoclonal antibodies (mAbs) that recognize FLuc. The mAbs, DSHB-LUC-2, DSHB-LUC-3, DSHB-LUC-9, DSHB-LUC-16, and DSHB-LUC-24, were generated by immunizing mice with purified 6xHIS-tagged FLuc (6xHis-FLuc) in suspension with an adjuvant. All five were validated by dot blots. Four of the mAbs provided strong signals in western blot analysis, and one a weak signal. All five were validated for immunostaining in fixed cell culture...
February 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Azam Hemmati, Hadi Hassannia, Saeideh Milani, Reza Hadavi, Fatemeh Ghaemimanesh, Hodjattallah Rabbani
Receptor tyrosine kinase ROR1 has been introduced as an interesting prognostic cancer marker in histopathology. The aim of this study was to produce a polyclonal antibody (PAb) against recombinant human ROR1 protein to be used as a tool for investigation of ROR1 expression in human cancer tissue blocks. The extracellular part of human ROR1 recombinant protein was expressed using pET-28b(+) plasmid in Escherichia coli Bl21(DE3) host. The recombinant ROR1, as a candidate immunogen, was purified and injected to a New Zealand rabbit...
February 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Tomohiro Osaki, Takeshi Nakanishi, Motoshi Aoki, Takahiro Omizu, Daisuke Nishiura, Masaya Kitamura
Tumor-targeted antibody-cytokine fusion proteins, called immunocytokines, are expected to be a useful platform for the development of effective antitumor therapeutic agents; however, their design and cost-efficient production remain as challenges. In this study, we constructed an antibody-cytokine fusion protein (Ia1-TNFα) comprising the single-domain antibody Ia1, which targets epidermal growth factor receptor (EGFR) overexpressed in epithelial tumors and a tumor necrosis factor α (TNFα) domain, which has antitumor activity...
February 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Amin Ramezani, Abbas Ghaderi
Targeted therapy using monoclonal antibodies (mAbs) against epidermal growth factor receptor-2 (ErbB2) has been utilized for the treatment of breast cancer, recently. Pertuzumab, one of the anti-ErbB2 mAbs, was approved by FDA in 2012 for the treatment of metastatic breast cancer. The aim of this study was to produce biosimilar version of pertuzumab in Chinese hamster ovary (CHO)-S cell line, and compare its ErbB2-binding and biological activities, with commercial drug, Perjeta. To this end, a dihydrofolate reductase (DHFR)-based strategy was used to produce a CHO-S stable cell pool capable of producing high levels of pertuzumab...
February 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Pei-Pei Meng, Zhe Li, Sheng-Yu Wang, Wen-Wen Zhou, Malik Samiullah, Na Chen, Fang-Hong Luo, Ting Wu, Jiang-Hua Yan
Class three semaphorins were originally identified as mediators of axon guidance, which repelled axons and collapsed growth cones. As a member of class three semaphorins, semaphorin3F (Sema3F) has been found to have similar effects on tumor cells and endothelial cells and also is implicated in the signaling of tumor metastasis by forming a complex with neuropilins and plexins. In this study, our laboratory produced a monoclonal antibody against the C-terminal domain of Sema3F (Sema3Fc mAb) using the hybridoma method, expecting to explore the potential role of the antibody and its application in the detection of Sema3F...
February 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Tomoto Suzuki, Yuya Takakubo, Hiroharu Oki, Xing Liu, Ryusuke Honma, Yasushi Naganuma, Stuart B Goodman, Mika K Kaneko, Yukinari Kato, Michiaki Takagi
Podoplanin (PDPN) is a transmembrane sialoglycoprotein, which is expressed in several normal tissues and malignant tumors. Although PDPN expression in rheumatoid arthritis (RA) has been reported, the role of PDPN in RA and other arthritic conditions has not been fully elucidated. In this study, we examined PDPN expression in inflammatory synovial tissues using an anti-human PDPN (hPDPN) monoclonal antibody (mAb) panel to select the most useful one for evaluation of synovitis. Synovial tissue samples were obtained from 11 RA patients and 9 osteoarthritis (OA) patients undergoing joint surgery...
February 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
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