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Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

Yoshikazu Furusawa, Shunsuke Itai, Shinji Yamada, Mika K Kaneko, Yukinari Kato
Telomerase reverse transcriptase (TERT) is the main catalytic subunit of telomerase-associated protein machinery. Upregulation of TERT at the transcriptional level results in immortal cell phenotype associated with cancer. To date we have developed eight anti-TERT monoclonal antibodies (mAbs) (TMab-4, TMab-5, TMab-6, TMab-7, TMab-9, TMab-10, TMab-11, and TMab-12) by immunizing mice with synthetic peptides (302 -QHHAGPPSTSRPPRPWDTPC-321 ) of TERT. We further characterized those epitopes using enzyme-linked immunosorbent assay, and here we discuss the critical epitope of an anti-TERT mAb, which is applicable for immunohistochemical analysis...
July 13, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Robert Hnasko, Alice V Lin, Larry Stanker, Jeffery McGarvey
Macrophage-conditioned medium (MCM) is an important cell culture supplement used to support the survival and growth of newly fused hybridoma cells. The use of macrophage cells, as a part of hybridoma technology, has proven to be an effective and inexpensive source of growth factors that promote the early survival and growth of hybridoma cells. Despite the widespread use of MCM as a hybridoma culture supplement, there is limited guidance and standardization for MCM production to achieve optimal hybridoma support...
June 14, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Ivan M Petyaev, Naylia A Zigangirova, Valeriy V Tsibezov, Elena Y Morgunova, Natalia E Bondareva, Nigel H Kyle, Yuriy K Bashmakov
Incubation of B10.MLM cells, a cell line of alveolar macrophages, with lycopene, a carotenoid, leads to an increase of lycopene content in their microsomal fraction. That increase was higher and developed faster when the cells were incubated with immune complexes formed by lycopene and mAb 6B9 (L-6B9 mAb), a monoclonal hapten-specific antibody raised against lycopene, as compared with dimethyl sulfoxide (DMSO)-dissolved lycopene (DMSO-L). Moreover, incubation of B10.MLM cells with L-6B9 mAb complexes was accompanied by more efficient accumulation of lipid droplets in the cultured cells and more significant inhibition of mRNA for 3-hydroxy-3-methylglutaryl-coenzyme (HMG-CoA) reductase, a rate-limiting enzyme of cholesterol biosynthesis known to be targeted by lycopene...
June 14, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Ivan M Petyaev, Naylia A Zigangirova, Dmitry Pristensky, Marina Chernyshova, Valeriy V Tsibezov, Natalya E Chalyk, Elena Y Morgunova, Nigel H Kyle, Yuriy K Bashmakov
Circulating lycopene level is negatively associated with the prevalence of cardiovascular disease, cancers (prostate and breast), type 2 diabetes mellitus, and aging. Traditionally, lycopene is measured in biological specimens by a combination of high-performance liquid chromatography (HPLC) and mass spectrometry methods. Moreover, as we recently reported, tissue/cell lycopene depositions can be observed by the immunohistochemistry method with a newly developed monoclonal antibody (mAb) against lycopene. A main objective of this study is to evaluate the performance of a new noninvasive immunofluorescence (IF) lycopene quantification skin test with mAbs against lycopene versus HPLC lycopene assay of serum lycopene in volunteers subjected to lycopene supplementation and represents its novelty...
June 14, 2018: 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
Anastas Pashov, Cinthia Violeta Hernandez Puente, Saddam Mohammed Ibrahim, Behjatolah Monzavi-Karbassi, Issam Makhoul, Thomas Kieber-Emmons
Evolutionary theories are necessarily invoked for understanding cancer development at the level of species, at the level of cells and tissues, and for developing effective therapies. It is crucial to view cancer in a Darwinian light, where the differential survival of individual cells is based on heritable variations. In the process of this somatic evolution, multicellularity controls are overridden by cancer cells, which become increasingly autonomous. Ecological epigenetics also helps understand how rogue cells that have basically the same DNA as their normal cell counterpart overcome the tissue homeostasis...
June 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
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June 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Xiaoman Wang, Yongzhen Zhao, Ling Li, Ziqiang Cheng, Guihua Wang
TRIM62 is a member of the tripartite interaction motif (TRIM) family and exerts crucial roles in innate immune response and cancer. To investigate the relationship between its distribution and avian retrovirus replication, in the present study, a monoclonal antibody (mAb) against chicken TRIM62 was generated. The open reading frame of chicken TRIM62 was amplified by polymerase chain reaction (PCR) and inserted into the expression vector pET-28a. The recombinant expression vectors were transformed into Escherichia coli BL21 (DE3)...
June 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Daisuke Oikawa, Masayuki Shiota, Fuminori Tokunaga, Hideki Wanibuchi
DAZ interacting zinc finger 3 (DZIP3), an RNA-binding RING-type ubiquitin ligase, has been reported to be involved in multiple physiological functions, including the regulation of chemokine- or estradiol-induced gene expression, self-renewal, and maintaining pluripotency in mouse embryonic stem cells. However, the precise cellular functions of DZIP3 remain elusive. In this study, we report the establishment of DZIP3-specific monoclonal antibodies (MAbs), using the rat medial iliac lymph node method. In immunoblotting analyses, our antibodies detected endogenous human and mouse DZIP3...
June 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, 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
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
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...
April 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 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 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 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 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...
April 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
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