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https://www.readbyqxmd.com/read/28424886/detection-of-koi-herpesvirus-khv-using-a-monoclonal-antibody-against-cyprinus-carpio-igm
#1
Yingying Li, Shucheng Zheng, Qing Wang, Sven M Bergmann, Weiwei Zeng, Yingying Wang, Chun Liu, Cunbin Shi
Koi herpesvirus disease (KHVD) is associated with high mortality in both common carp and koi carp (Cyprinus carpio L.) worldwide. The indirect detection of fish viruses based on the identification of antibodies has emerged as a practical and reliable means of diagnosis. Thus, it is important to create monoclonal antibodies (MAbs) against carp IgM. By using hybridoma-monoclonal antibody technology, one hybridoma cell line secreting MAbs against IgM from carp was established. In western blot analysis, the secreted MAb from cell line A5-E10 recognized the heavy chain of IgM from common carp or koi but did not react with immunoglobulins from three different fish species: grass carp (Ctenopharyngodon idella), tilapia (Oreochromis mossambicus) and Mandarin fish (Siniperca chuatsi)...
April 19, 2017: Archives of Virology
https://www.readbyqxmd.com/read/28421663/cloning-and-plant-based-production-of-antibody-mc10e7-for-a-lateral-flow-immunoassay-to-detect-4-arginine-microcystin-in-fresh-water
#2
Stanislav Melnik, A-C Neumann, R Karongo, S Dirndorfer, Martin Stübler, Verena Ibl, R Niessner, Dietmar Knopp, Eva Stoger
Antibody MC10E7 is one of a small number of monoclonal antibodies that bind specifically to [Arg4]-microcystins and it can be used to survey natural water sources and food samples for algal toxin contamination. However, the development of sensitive immunoassays in different test formats, particularly user-friendly tests for on-site analysis, requires a sensitive but also cost-effective antibody. The original version of MC10E7 was derived from a murine hybridoma, but we determined the sequence of the variable regions using the peptide mass-assisted cloning strategy and expressed a scFv (single-chain variable fragment) format of this antibody in yeast and a chimeric full size version in leaves of Nicotiana tabacum and N...
April 19, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28417626/production-and-characterization-of-monoclonal-antibody-against-recombinant-virus-coat-protein-cp42
#3
Naeimeh Shibaei, Jafar Majidi, Khadijeh Razavi, Ali Asghar Karkhane, Nemat Sokhandan-Bashir, Leili Aghebati-Maleki
There are many studies related to the production of a ELISA kit for diagnosing virus infections. However, production of most kits depends on purification of whole virus particles, which involves the use of costly equipment and reagents. The purpose of this study was to check out if the anti-CP42 antibodies could be used as a diagnostic assay for detection of Grapevine fanleaf Virus (GFLV). In this study, recombinant GFLV coat protein gene related to selected antigenic determinants was inserted into pET-28a bacterial expression vector and the construct (pET-28a CP42) was cloned into E...
February 2017: Iranian Journal of Allergy, Asthma, and Immunology
https://www.readbyqxmd.com/read/28414197/engineering-of-a-self-adjuvanted-itep-delivered-ctl-vaccine
#4
Shuyun Dong, Tiefeng Xu, Peng Wang, Peng Zhao, Mingnan Chen
Cytotoxic T lymphocyte (CTL) epitope peptide-based vaccines are widely used in cancer and infectious disease therapy. We previously generated an immune-tolerant elastin-like polypeptides (iTEPs)-based carrier to deliver a peptide CTL vaccine and enhance the efficiency of the vaccine. To further optimize the vaccine carrier, we intended to potentiate its function by designing an iTEP-based carrier that was able to deliver adjuvant and a vaccine epitope as one molecule. Thus, we fused a 9-mer H100, a peptide derived from the high-mobility group box 1 protein (HMGB1) that could induce activation of dendritic cells (DCs), with an iTEP polymer to generate a new iTEP polymer named H100-iTEP...
April 17, 2017: Acta Pharmacologica Sinica
https://www.readbyqxmd.com/read/28414151/targeted-antigen-delivery-to-dendritic-cell-via-functionalized-alginate-nanoparticles-for-cancer-immunotherapy
#5
Chuangnian Zhang, Gaona Shi, Ju Zhang, Huijun Song, Jinfeng Niu, Shengbin Shi, Pingsheng Huang, Yanming Wang, Weiwei Wang, Chen Li, Deling Kong
The purpose of the present study was to identify an "easy-to-adopt" strategy to enhance immune responses using functionalized alginate (ALG) nanoparticles (MAN-ALG/ALG=OVA NPs), which were prepared by CaCl2 cross-linking of two different types of ALG. The mannose (MAN) modified ALG (MAN-ALG) was used for dendritic cell targeting. The other component, composed of ovalbumin (OVA), a model antigen, is conjugated to ALG (ALG=OVA) via pH sensitive Schiff base bond. Grafting of alginate was demonstrated by FT-IR and (1)H NMR, while the morphological structure, particle size, Zeta potential of MAN-ALG/ALG=OVA NPs were measured using TEM and DLS...
April 13, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28414024/novel-high-throughput-cell-based-hybridoma-screening-methodology-using-the-celigo-image-cytometer
#6
Haohai Zhang, Leo Li-Ying Chan, William Rice, Nasim Kassam, Maria Serena Longhi, Haitao Zhao, Simon C Robson, Wenda Gao, Yan Wu
Hybridoma screening is a critical step for antibody discovery, which necessitates prompt identification of potential clones from hundreds to thousands of hybridoma cultures against the desired immunogen. Technical issues associated with ELISA- and flow cytometry-based screening limit accuracy and diminish high-throughput capability, increasing time and cost. Conventional ELISA screening with coated antigen is also impractical for difficult-to-express hydrophobic membrane antigens or multi-chain protein complexes...
April 14, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28412548/internal-image-anti-idiotypic-antibody-a-new-strategy-for-the-development-a-new-category-of-prolactin-receptor-prlr-antagonist
#7
Hainan Lan, Pan Hong, Ruonan Li, Suo L, Shan Anshan, Steven Li, Xin Zheng
Over the past decades, a number of prolactin receptor (PRLR) antagonists have been developed, which can be divided into two categories, PRLR analogue and anti-PRLR antibody. However, until now, there have been no commercially available PRLR antagonists. Here, we described a new approach for the preparation of PRLR antagonist, namely internal image anti-idiotypic antibody strategy. The hybridoma technique was used to generate anti-idiotypic antibodies to PRL. Competitive ELISA, competitive receptor-binding analysis and immunofluorescence assay (IFA) were then used to screen and characterize anti-idiotypic antibodies to PRL...
April 13, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28409665/monoclonal-antibody-against-premembrane-viral-protein-of-avian-tembusu-virus
#8
Xiaozhi Huang, Tingting Shi, Kaikun Mo, Dandan Wang, Xing Peng, Min Liao, Jiyong Zhou
Premembrane (prM) is a viral protein of flavivirus, which is important for the generation of infectious virion and for virus infection to the host. However, the biological properties and function of the prM of Avian Tembusu virus (ATMUV) have scarcely been studied to date. Monoclonal antibodies (mAbs) are a powerful tool for functional analysis of viral protein. To produce a mAb against prM protein of ATMUV, the prM gene sequence was amplified by reverse transcription polymerase chain reaction (RT-PCR) and cloned into the prokaryotic expression vector pET-28a (+)...
April 14, 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28402743/generation-and-application-of-monoclonal-antibody-against-lycopene
#9
Valeriy V Tsibezov, Yuriy K Bashmakov, Dmitry V Pristenskiy, Naylia A Zigangirova, Ludmila V Kostina, Natalya E Chalyk, Alexey Y Kozlov, Elena Y Morgunova, Marina P Chernyshova, Marina V Lozbiakova, Nigel H Kyle, Ivan M Petyaev
A monoclonal antibody (Mab) against lycopene was developed from hybridoma clones obtained from BALB/c mice immunized with trans-isomer of lycopene (t-lycopene, t-LC) conjugated with colloidal gold particles. An alternating immunization schedule which included injection of both formulations of immunogen (without and with Freund's adjuvant) was most effective in the elucidation of a measurable immune response to the t-Lycopene conjugate. Selected hybridoma clones were able to produce an Mab positive in competition assay...
April 12, 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28400756/antibody-engineering-for-pursuing-a-healthier-future
#10
REVIEW
Abdullah F U H Saeed, Rongzhi Wang, Sumei Ling, Shihua Wang
Since the development of antibody-production techniques, a number of immunoglobulins have been developed on a large scale using conventional methods. Hybridoma technology opened a new horizon in the production of antibodies against target antigens of infectious pathogens, malignant diseases including autoimmune disorders, and numerous potent toxins. However, these clinical humanized or chimeric murine antibodies have several limitations and complexities. Therefore, to overcome these difficulties, recent advances in genetic engineering techniques and phage display technique have allowed the production of highly specific recombinant antibodies...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28395728/-preparation-and-characterization-of-monoclonal-antibody-against-gp120-v1-v2-domain-of-hiv-1-subtype-crf01_ae
#11
Ying Chu, Xiangdan Gong, Jianwei Gao, Airong Su, Deyan Chen, Hongyong Song, Xilin Wu, Zhiwei Wu
Objective To express the V1/V2 domain of HIV-1 subtype CRF01_AE gp120 in eukaryotic cells, and then prepare its monoclonal antibody (mAb) and identify its antigen reactivity. Methods Eukaryotic expression vector of pTriEx-3-V1/V2CNE55 was constructed and transiently transfected into HEK293F cell line. The V1/V2-His chimera protein was purified and injected into BALB/c mice. After the fusion between spleen cells of immunized BALB/c mice and myeloma cells SP 2/0, ELISA was used for screening the positive hybridoma cell clones against the V1/V2 recombinant protein...
April 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28391164/antibody-based-targeting-of-cd24-enhances-antitumor-effect-of-cetuximab-via-attenuating-phosphorylation-of-src-stat3
#12
Zhiguo Chen, Tong Wang, Xiaojie Tu, Wei Xie, Hua He, Min Wang, Juan Zhang
Epidermal growth factor receptor (EGFR) is a cell-surface receptor for some extracellular protein ligands relating to cancers and has been recognized as a key target for tumor therapy. Cetuximab, a chimerical monoclonal EGFR IgG1 antibody, is used for the treatment of various malignancies. However, recent clinical trials reported that the anti-tumor effect of cetuximab is still controversial. Cluster of differentiation 24 (CD24) is a tumor-associated antigen correlating with poor prognosis and regulating the activity of Src/STAT3 in multiple cancers...
April 5, 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/28369682/characterization-and-functional-analysis-of-mouse-semi-invariant-natural-t-cells
#13
Amrendra Kumar, Jelena S Bezbradica, Aleksandar K Stanic, Sebastian Joyce
Semi-invariant natural killer T (iNKT) cells are CD1d-restricted innate-like lymphocytes that recognize lipid agonists. Activated iNKT cells have immunoregulatory properties. Human and mouse iNKT cell functions elicited by different glycolipid agonists are highly conserved, making the mouse an excellent animal model for understanding iNKT cell biology in vivo. This unit describes basic methods for the characterization and quantification (see Basic Protocol 1) and functional analysis of mouse iNKT cells in vivo or in vitro...
April 3, 2017: Current Protocols in Immunology
https://www.readbyqxmd.com/read/28359942/production-characterization-and-application-of-monoclonal-antibody-against-immunoglobulin-d-heavy-chain-of-flounder-paralichthys-olivaceus
#14
Xiaoqian Tang, Fuguo Liu, Xiuzhen Sheng, Jing Xing, Wenbin Zhan
Immunoglobulin D (IgD) is considered to be an enigmatic Ig molecule because of the lack understanding of its immunological functions. In the present study, a partial δ region of the flounder IgD was recombinantly expressed, purified and used as an immunogen to produce monoclonal antibodies (MAbs) against the H chain of flounder IgD. After fusion, a total of 97 hybridomas were generated and observed under an inverted microscope One of the hybridomas, designated 5G7, gave strong positive results in an indirect enzyme-linked immunosorbent assay (ELISA) and was cloned and subcloned by limiting dilution...
May 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/28336958/from-rabbit-antibody-repertoires-to-rabbit-monoclonal-antibodies
#15
REVIEW
Justus Weber, Haiyong Peng, Christoph Rader
In this review, we explain why and how rabbit monoclonal antibodies have become outstanding reagents for laboratory research and increasingly for diagnostic and therapeutic applications. Starting with the unique ontogeny of rabbit B cells that affords highly distinctive antibody repertoires rich in in vivo pruned binders of high diversity, affinity and specificity, we describe the generation of rabbit monoclonal antibodies by hybridoma technology, phage display and alternative methods, along with an account of successful humanization strategies...
March 24, 2017: Experimental & Molecular Medicine
https://www.readbyqxmd.com/read/28304351/the-preparation-and-identification-of-a-monoclonal-antibody-against-citrinin-and-the-development-of-detection-via-indirect-competitive-elisa
#16
Shimuye Kalayu Yirga, Sumei Ling, Yanling Yang, Jun Yuan, Shihua Wang
Citrinin (CTN) is a hepato-nephrotoxic mycotoxin produced by fungi genera of Aspergillus, Monauscus, and Penicillium. CTN contaminates grains, fruits, juices and vegetables, and causes various toxic effects to humans and animals. It has small molecular weight, which is non-immunogenic to animals. Thus, CTN was conjugated to bovine serum albumin (BSA) and ovalbumin (OVA), respectively, by amide bonds using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). Mice were immunized with CTN-BSA conjugates, and spleen cells of the immunized mice were fused with Sp2/0 myeloma cells to obtain 21H27 hybriodoma cell...
March 17, 2017: Toxins
https://www.readbyqxmd.com/read/28296410/a-light-responsive-nanoparticle-based-delivery-system-using-pheophorbide-a-graft-polyethylenimine-for-dendritic-cell-based-cancer-immunotherapy
#17
Chuangnian Zhang, Ju Zhang, Gaona Shi, Huijuan Song, Shengbin Shi, Xiuyuan Zhang, Pingsheng Huang, Zhihong Wang, Weiwei Wang, Chun Wang, Deling Kong, Chen Li
In this study, the photochemical internalization (PCI) technique was adopted in a nanoparticle-based antigen delivery system to enhance antigen-specific CD8(+) T cell immune response for cancer immunotherapy. Pheophorbide A, a hydrophobic photosensitizer, grafted with polyethylenimine (PheoA-PEI) with endosome escape activity and near-infrared imaging capability was prepared. A model antigen ovalbumin (OVA) was then complexed with PheoA-PEI to form PheoA-PEI/OVA nanoparticles (PheoA-PEI/OVA NPs) that are responsive to light...
March 28, 2017: Molecular Pharmaceutics
https://www.readbyqxmd.com/read/28293253/can-anti-bothropstoxin-i-antibodies-discriminate-between-bothrops-jararaca-and-bothrops-jararacussu-venoms
#18
Ricardo Teixeira Araujo, Carlos Corrêa-Netto, Leonora Brazil-Más, Caio Raony Farina Silveira, Irene Fernandes, Russolina Benedeta Zingali
BACKGROUND: Snakes of the genus Bothrops, popularly known as pit vipers, are responsible for most cases of snakebite in Brazil. Within this genus, Bothrops jararacussu and B. jararaca deserve special attention due to the severity of their bites and for inhabiting densely populated areas. Regarding the treatment of snakebites by Bothrops jararacussu, questions have been raised about the effectiveness of the specific bothropic antivenom in neutralizing myotoxic effects; however, there are no accurate data for humans...
2017: Journal of Venomous Animals and Toxins Including Tropical Diseases
https://www.readbyqxmd.com/read/28286781/generation-of-monoclonal-antibodies-against-immunoglobulin-proteins-of-the-domestic-ferret-mustela-putorius-furo
#19
Greg A Kirchenbaum, Ted M Ross
The domestic ferret (Mustela putorius furo) serves as an animal model for the study of several viruses that cause human disease, most notably influenza. Despite the importance of this animal model, characterization of the immune response by flow cytometry (FCM) is severely hampered due to the limited number of commercially available reagents. To begin to address this unmet need and to facilitate more in-depth study of ferret B cells including the identification of antibody-secreting cells, eight unique murine monoclonal antibodies (mAb) with specificity for ferret immunoglobulin (Ig) were generated using conventional B cell hybridoma technology...
2017: Journal of Immunology Research
https://www.readbyqxmd.com/read/28282494/hapten-specific-single-cell-selection-of-hybridoma-clones-by-fluorescence-activated-cell-sorting-for-the-generation-of-monoclonal-antibodies
#20
Martin Dippong, Peter Carl, Christine Lenz, Jörg A Schenk, Katrin Hoffmann, Timm Schwaar, Rudolf J Schneider, Maren Kuhne
The conventional hybridoma screening and subcloning process is generally considered to be one of the most critical steps in hapten-specific antibody production. It is time-consuming, monoclonality is not guaranteed, and the number of clones that can be screened is limited. Our approach employs a novel hapten-specific labeling technique of hybridoma cells. This allows for fluorescence-activated cell sorting (FACS) and single-cell deposition and thereby eliminates the above-mentioned problems. A two-step staining approach is used to detect antigen specificity and antibody expression: in order to detect antigen specificity, hybridoma cells are incubated with a hapten-horseradish peroxidase conjugate (hapten-HRP), which is subsequently incubated with a fluorophore-labeled polyclonal anti-peroxidase antibody (anti-HRP-Alexa Fluor 488)...
March 22, 2017: Analytical Chemistry
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