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https://www.readbyqxmd.com/read/28719245/production-of-a-mouse-monoclonal-antibody-against-mortalin-by-whole-cell-immunization
#1
Marzieh Rezaei, Abbas Ghaderi
Pancreatic carcinoma is the fourth leading cause of cancer death and is characterized by early invasion and metastasis. Advances in molecular biology directed new strategies in targeted therapy using monoclonal antibodies. To identify new biomarkers, we generated a panel of monoclonal antibodies against the newly established cell line, Faraz-ICR, from a patient with acinar cell carcinoma. After immunization of BALB/c female mice with Faraz-ICR cell line and fusion of splenocytes with SP2/0 myeloma cell line, high reactive hybridoma producing antibodies to Faraz-ICR were detected using enzyme-linked immunosorbent assay, immunofluorescence staining and flow cytometry...
July 18, 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28713817/generation-of-monoclonal-antibodies-against-ag85a-antigen-of-mycobacterium-tuberculosis-and-application-in-a-competitive-elisa-for-serodiagnosis-of-bovine-tuberculosis
#2
Zhengzhong Xu, Ting Hu, Aihong Xia, Xin Li, Ze Liu, Jingjing Min, Jingjing He, Chuang Meng, Yuelan Yin, Xiang Chen, Xinan Jiao
The Ag85 complex functions as the main secretory protein of Mycobacterium tuberculosis (M. tuberculosis) and BCG. This complex is composed of the proteins, Ag85A, Ag85B, and Ag85C, with Ag85A thought to play the largest role within the complex. However, the lack of commercially available monoclonal antibodies (mAbs) against Ag85A still hinders the biological and applicative research on this protein. In this study, we developed and identified anti-Ag85A mAbs, and five hybridoma cells were established. Using the indirect immunofluorescence test, we found that two anti-Ag85A mAbs did not cross-react with Ag85B and/or Ag85C...
2017: Frontiers in Veterinary Science
https://www.readbyqxmd.com/read/28712409/-preparation-and-identification-of-monoclonal-antibodies-against-chicken-interleukin-4
#3
Hua Dai, Junhua Chen, Haixia Zhang, Ying Shen, Xiang Chen, Xin'an Jiao
Objective To prepare and characterize the monoclonal antibodies (mAbs) against chicken interleuikin-4 (ChIL-4). Methods Using lymphocyte hybridoma technique, 8-week-old BALB/c mice were immunized with purified recombinant ChIL-4 (rChIL-4) with GST tag (GST-ChIL-4), and rChIL-4 with histidine tag (His-ChIL-4) was utilized to set up an indirect ELISA to screen positive hybridomas. Limited dilution methods were used to generate hybridoma cell lines stably secreting mAbs against ChIL-4. The specificity of the mAbs was identified by indirect ELISA, immunofluorescent assay (IFA) and Western blot...
July 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28692735/an-anti-vegf-b-antibody-fragment-induces-regression-of-pre-existing-blood-vessels-in-the-rat-cornea
#4
Yazad D Irani, Pierre D Scotney, Sonja Klebe, Lauren A Mortimer, Andrew D Nash, Keryn A Williams
Purpose: We tested the ability of an antibody fragment with specificity for vascular endothelial growth factor-B (VEGF-B) to regress nascent and established corneal blood vessels in the rat. Methods: A single chain variable antibody fragment (scFv) with specificity for VEGF-B was engineered from the 2H10 hybridoma. Binding to rat, mouse, and human VEGF-B was confirmed by surface plasmon resonance. Activity of the anti-VEGF-B scFv on developing and established corneal blood vessels was assessed following unilateral superficial cautery in male and female outbred Sprague Dawley rats...
July 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/28679076/development-of-a-tree-shrew-specific-interferon-gamma-assay
#5
Xuemei Zhang, Jingwen Xu, Zhongxiang Wu, Wenbing Zhu, Shaozhong Dong
Tree shrews (Tupaia belangeri) are small squirrel-like mammals closely related to primates. Due to their susceptibility to several human viruses, tree shrews have been proposed as potential animal models for the study of human viral infections. However, there are no standardized assays currently available for the detection of tree shrew-specific interferon (IFN)-γ, a major cytokine secreted during the antiviral immune response. Herein, we developed a novel enzyme-linked immunosorbent assay (ELISA) for the quantification of IFN-γ in tree shrew serum samples...
July 5, 2017: Journal of Immunoassay & Immunochemistry
https://www.readbyqxmd.com/read/28677761/expression-and-purification-of-a-major-allergen-pla-a-1-from-platanus-acerifolia-pollen-and-the-preparation-of-its-monoclonal-antibody
#6
Wei-Wei Ni, Wen Huang, De-Qin Wu, Yan-Jun Zhou, Chun-Mei Ji, Meng-Da Cao, Miao Guo, Jin-Lu Sun, Ji-Fu Wei
Platanus acerifolia pollen is considered an important source of airborne allergens in numerous cities. Pla a 1 is a major allergen from P. acerifolia pollen. The present study aimed to express and purify Pla a 1, and to prepare its monoclonal antibody. In the present study, the Pla a 1 gene was subcloned into a pET‑28a vector and transformed into the ArcticExpress™ (DE3) RP Escherichia coli host strain. The purified Pla a 1 was then used to immunize BALB/c mice. When serum detection was positive, spleen cells were isolated from the mice and fused with SP2/0 myeloma cells at a ratio of 10:1...
September 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28674295/development-of-a-rapid-identification-method-for-a-variety-of-antibody-candidates-using-high-throughput-sequencing
#7
Yuji Ito
 As an alternative to hybridoma technology, the antibody phage library system can also be used for antibody selection. This method enables the isolation of antigen-specific binders through an in vitro selection process known as biopanning. While it has several advantages, such as an avoidance of animal immunization, the phage cloning and screening steps of biopanning are time-consuming and problematic. Here, we introduce a novel biopanning method combined with high-throughput sequencing (HTS) using a next-generation sequencer (NGS) to save time and effort in antibody selection, and to increase the diversity of acquired antibody sequences...
2017: Yakugaku Zasshi: Journal of the Pharmaceutical Society of Japan
https://www.readbyqxmd.com/read/28672746/study-of-kallikrein-related-peptidase-6-klk6-and-its-complex-with-%C3%AE-1-antitrypsin-in-biological-fluids
#8
Dimitrios Korbakis, Antoninus Soosaipillai, Eleftherios P Diamandis
BACKGROUND: Human kallikrein-related peptidase 6 (KLK6) is a member of the kallikrein family of serine proteases. KLK6 is synthesized as a preproenzyme, mainly in tissues of the central nervous system (CNS), and secreted as an inactive precursor. Serum KLK6 is a biomarker of unfavorable prognosis for ovarian cancer, but its sensitivity for early detection is relatively low. Differential glycosylation of KLK6 has been identified in ascites fluid obtained from ovarian cancer patients, suggesting the presence of unique KLK6 isoforms in biological samples...
June 24, 2017: Clinical Chemistry and Laboratory Medicine: CCLM
https://www.readbyqxmd.com/read/28662180/kai-1-and-kai-2-characterization-of-these-dog-erythrocyte-antigens-by-monoclonal-antibodies
#9
Jae Ho Lee, Urs Giger, Hee Young Kim
Dog Erythrocyte Antigens (DEA) have thus far been found by sensitizing dogs with canine allogeneic blood and are clinically important regarding blood transfusion incompatibilities, but remain poorly defined. The goals of this study were to discover and characterize two DEAs, named as Kai 1 and Kai 2. The monoclonal antibodies were produced by mouse hybridoma techniques and examined by ELISA isotyping, immunoblotting, and affinity chromatography. Canine blood samples were typed and the development of alloantibodies was examined in transfused dogs...
2017: PloS One
https://www.readbyqxmd.com/read/28640955/in-solution-antibody-harvesting-with-a-plant-produced-hydrophobin-protein-a-fusion
#10
Katri Kurppa, Lauri Reuter, Anneli Ritala, Markus Linder, Jussi Joensuu
Purification is a bottleneck and a major cost factor in the production of antibodies. We set out to engineer a bi-functional fusion protein from two building blocks, Protein A and a hydrophobin, aiming at low-cost and scalable antibody capturing in solutions. Immunoglobulin-binding Protein A is widely used in affinity-based purification. The hydrophobin fusion tag, on the other hand, has been shown to enable purification by two-phase separation. Protein A was fused to two different hydrophobin tags, HFBI or II, and expressed transiently in Nicotiana benthamiana...
June 22, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28632753/mait-cells-launch-a-rapid-robust-and-distinct-hyperinflammatory-response-to-bacterial-superantigens-and-quickly-acquire-an-anergic-phenotype-that-impedes-their-cognate-antimicrobial-function-defining-a-novel-mechanism-of-superantigen-induced-immunopathology
#11
Christopher R Shaler, Joshua Choi, Patrick T Rudak, Arash Memarnejadian, Peter A Szabo, Mauro E Tun-Abraham, Jamie Rossjohn, Alexandra J Corbett, James McCluskey, John K McCormick, Olivier Lantz, Roberto Hernandez-Alejandro, S M Mansour Haeryfar
Superantigens (SAgs) are potent exotoxins secreted by Staphylococcus aureus and Streptococcus pyogenes. They target a large fraction of T cell pools to set in motion a "cytokine storm" with severe and sometimes life-threatening consequences typically encountered in toxic shock syndrome (TSS). Given the rapidity with which TSS develops, designing timely and truly targeted therapies for this syndrome requires identification of key mediators of the cytokine storm's initial wave. Equally important, early host responses to SAgs can be accompanied or followed by a state of immunosuppression, which in turn jeopardizes the host's ability to combat and clear infections...
June 2017: PLoS Biology
https://www.readbyqxmd.com/read/28618148/mechanisms-underlying-induction-of-allergic-sensitization-by-pru-p-3
#12
L Tordesillas, N Cubells-Baeza, C Gómez-Casado, C Berin, V Esteban, W Barcik, L O'Mahony, C Ramirez, L F Pacios, M Garrido-Arandia, A Díaz-Perales
BACKGROUND: Recently, the nature of the lipid-ligand of Pru p 3, one of the most common plant food allergens in southern Europe, has been identified as a derivative of the alkaloid camptothecin bound to phytosphingosine. However, the origin of its immunological activity is still unknown. OBJECTIVE: We sought to evaluate the role of the Pru p 3 lipid-ligand in the immunogenic activity of Pru p 3. METHODS: In vitro cultures of different cell types (monocyte-derived dendritic cells [moDCs], PBMCs [peripheral blood mononuclear cells] and epithelial and iNKT-hybridoma cell lines) have been used to determine the immunological capacity of the ligand, by measuring cell proliferation, maturation markers and cytokine production...
June 15, 2017: Clinical and Experimental Allergy: Journal of the British Society for Allergy and Clinical Immunology
https://www.readbyqxmd.com/read/28592796/immunochemical-approach-for-monitoring-of-structural-transition-of-apoa-i-upon-hdl-formation-using-novel-monoclonal-antibodies
#13
Hitoshi Kimura, Shiho Mikawa, Chiharu Mizuguchi, Yuki Horie, Izumi Morita, Hiroyuki Oyama, Takashi Ohgita, Kazuchika Nishitsuji, Atsuko Takeuchi, Sissel Lund-Katz, Kenichi Akaji, Norihiro Kobayashi, Hiroyuki Saito
Apolipoprotein A-I (apoA-I) undergoes a large conformational reorganization during remodeling of high-density lipoprotein (HDL) particles. To detect structural transition of apoA-I upon HDL formation, we developed novel monoclonal antibodies (mAbs). Splenocytes from BALB/c mice immunized with a recombinant human apoA-I, with or without conjugation with keyhole limpet hemocyanin, were fused with P3/NS1/1-Ag4-1 myeloma cells. After the HAT-selection and cloning, we established nine hybridoma clones secreting anti-apoA-I mAbs in which four mAbs recognize epitopes on the N-terminal half of apoA-I while the other five mAbs recognize the central region...
June 7, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28592426/a-nonadjuvanted-igg2a-monoclonal-antibody-against-nucleosomes-elicits-potent-t-cell-dependent-idiotype-specific-igg1-responses-and-glomerular-igg1-igg2a-deposits-in-normal-mice
#14
Kristian Hannestad, Helge Scott
Idiotypes (Ids) are unique epitopes of Ab V regions and can trigger anti-Id immune responses, but immunization with several nonadjuvanted isologous IgG mAbs has induced tolerance to their Ids. We immunized non-lupus-prone mice with 11 allotype "a" of IgG2a (IgG2a(a)) and 4 IgG2c nonadjuvanted, isologous mAbs purified from serum-free medium. Of five IgG2a(a) mAbs with specificity for nucleosomes, the repeating histone-DNA subunit of chromatin, four elicited an IgG1 anti-mAb response and one mAb was nonimmunogenic...
June 7, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28589361/the-aid-cre-ert2-model-a-tool-for-monitoring-b-cell-immune-responses-and-generating-selective-hybridomas
#15
Simon Le Gallou, Takuya Nojima, Daisuke Kitamura, Jean-Claude Weill, Claude-Agnès Reynaud
Expression of activation-induced cytidine deaminase (AID) is the hallmark of B cells engaged in an immune response in germinal centers. We designed an inducible fate-mapping reporter mouse in which AID-expressing B cells could be timely and irreversibly marked, by knockin at the Aicda locus of a tamoxifen-inducible Cre recombinase. This mouse model allows notably for the long-term follow-up of memory B cells and plasma cells engaged in an immune response. We describe here a protocol to generate hybridomas from small memory subsets that can be easily traced and identified in this mouse line through Cre-activated fluorescent reporters...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28570826/specific-monoclonal-antibodies-recognizing-the-endogenous-chicken-high-mobility-group-box-1-protein
#16
Yurong Qu, Yuan Zhan, Shen Yang, Xusheng Qiu, Lei Tan, Yingjie Sun, Chunchun Meng, Cuiping Song, Shengqing Yu, Chan Ding
High mobility group box 1 (HMGB1) is a key member of the "danger associated molecular patterns" (DAMPs), which can localize in various compartments of the cell, and plays important roles in systemic inflammation. In the present study, monoclonal antibodies (MAbs) specifically against chicken HMGB1 were generated. The open reading frame of chicken HMGB1 was amplified by RT-PCR and cloned into the prokaryotic expression vector pET-28a to construct a recombinant plasmid pET-chHMGB1. The recombinant chicken HMGB1 protein was expressed in Escherichia coli Rosetta under IPTG induction and then purified by Ni-NTA Purification System...
June 1, 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28558726/development-and-application-of-triple-antibody-sandwich-enzyme-linked-immunosorbent-assays-for-begomovirus-detection-using-monoclonal-antibodies-against-tomato-yellow-leaf-curl-thailand-virus
#17
Channarong Seepiban, Saengsoon Charoenvilaisiri, Nuchnard Warin, Anjana Bhunchoth, Namthip Phironrit, Bencharong Phuangrat, Orawan Chatchawankanphanich, Supat Attathom, Oraprapai Gajanandana
BACKGROUND: Tomato yellow leaf curl Thailand virus, TYLCTHV, is a begomovirus that causes severe losses of tomato crops in Thailand as well as several countries in Southeast and East Asia. The development of monoclonal antibodies (MAbs) and serological methods for detecting TYLCTHV is essential for epidemiological studies and screening for virus-resistant cultivars. METHODS: The recombinant coat protein (CP) of TYLCTHV was expressed in Escherichia coli and used to generate MAbs against TYLCTHV through hybridoma technology...
May 30, 2017: Virology Journal
https://www.readbyqxmd.com/read/28557609/myeloma-derived-light-chain-paired-with-a-diagnostic-monoclonal-antibody-hinders-immunoassay-performance
#18
Bailin Tu, Bryan Tieman, Jeffrey Moore, You Pan, A Scott Muerhoff
Monoclonal antibodies are widely used as the capture and detection reagents in diagnostic immunoassays. In the past, myeloma fusion partners expressing endogenous heavy and/or light chains were often used to generate hybridoma cell lines. As a result, mixed populations of antibodies were produced that can cause inaccurate test results, poor antibody stability, and significant lot-to-lot variability. We describe one such scenario where the P3U1 (P3X63Ag8U.1) myeloma fusion partner was used in the generation of a hybridoma producing protein induced vitamin K absence/antagonist-II (PIVKA II) antibody...
June 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28545467/expression-of-deleted-atoxic-atypical-recombinant-beta2-toxin-in-a-baculovirus-system-and-production-of-polyclonal-and-monoclonal-antibodies
#19
Anna Serroni, Chiara Francesca Magistrali, Giovanni Pezzotti, Luca Bano, Martina Pellegrini, Giulio Severi, Chiara Di Pancrazio, Mirella Luciani, Manuela Tittarelli, Silvia Tofani, Antonio De Giuseppe
BACKGROUND: Clostridium perfringens is an important animal and human pathogen that can produce more than 16 different major and minor toxins. The beta-2 minor toxin (CPB2), comprising atypical and consensus variants, appears to be involved in both human and animal enterotoxaemia syndrome. The exact role of CPB2 in pathogenesis is poorly investigated, and its mechanism of action at the molecular level is still unknown because of the lack of specific reagents such as monoclonal antibodies against the CPB2 protein and/or the availability of a highly purified antigen...
May 25, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28531814/targeting-egfr-her2-heterodimerization-with-a-novel-anti-her2-domain-ii-iii-antibody
#20
Xiaojie Yu, Lingfei Wang, Yafeng Shen, Chao Wang, Yajun Zhang, Yanchun Meng, Yang Yang, Beibei Liang, Bo Zhou, Huajing Wang, Huafeng Wei, Changhai Lei, Shi Hu, Bohua Li
HER2, a ligand-free tyrosine kinase receptor of the HER family, is frequently overexpressed in breast cancer. The anti-HER2 antibody trastuzumab has shown significant clinical benefits in metastatic breast cancer. Despite the effectiveness of trastuzumab, its efficacy remains variable and often modest. Thus, there is an urgent need to improve ErbB2-targeting therapy. Here, we describe a novel anti-HER2 antibody, 7C3, which was developed using hybridoma technique. Structural analysis confirms that the epitope of this antibody is in domain II/III of HER2...
May 19, 2017: Molecular Immunology
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