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https://www.readbyqxmd.com/read/27894352/immunoassay-of-s-adenosylmethionine-and-s-adenosylhomocysteine-the-methylation-index-as-a-biomarker-for-disease-and-health-status
#1
Xiujuan Hao, Yan Huang, Ming Qiu, Chunlin Yin, Huiming Ren, Hongjie Gan, Huijun Li, Yaxia Zhou, Jiazhi Xia, Wenting Li, Lijuan Guo, Isaac A Angres
BACKGROUND: S-Adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) are relevant to a variety of diseases. Previous reports that quantified SAM and SAH were based on HPLC or LC-MS/MS. No antibody against SAM has been generated, and the antibody against SAH cannot be used with blood samples. Immunoassays have not been used to measure SAM and SAH. In this study, ELISA was used to measure blood SAM and SAH levels. RESULTS: Specific antibodies against SAM were produced for the first time using a stable analog as the antigen...
November 28, 2016: BMC Research Notes
https://www.readbyqxmd.com/read/27888901/-monoclonal-antibodies-against-pcsk9-from-bench-to-clinic
#2
Carlos Guijarro Herraiz
Antibodies are glycoproteins with high specificity binding to multiple antigens due to the large number of structural conformations of the variable chains. Hybridoma technology (fusion of myeloma cells with immunoglobulin-producing lymphocytes) has allowed the synthesis of large quantities of unique antibodies (monoclonal [mAb]). mAbs were initially murine. Subsequently, chimeric mAbs were developed, followed by humanized mAbs and finally human mAbs. The high selectivity and good tolerance of human mAbs allows their therapeutic administration to block specific exogenous or endogenous molecules...
May 2016: Clínica e Investigación en Arteriosclerosis
https://www.readbyqxmd.com/read/27876044/generation-of-monoclonal-antibodies-against-native-viral-proteins-using-antigen-expressing-mammalian-cells-for-mouse-immunization
#3
Natalie Rose, Carlos Augusto Pinho-Nascimento, Alessia Ruggieri, Paola Favuzza, Marco Tamborrini, Hanna Roth, Marcia Terezinha Baroni de Moraes, Hugues Matile, Thomas Jänisch, Gerd Pluschke, Katharina Röltgen
BACKGROUND: Due to their rising incidence and progressive geographical spread, infections with mosquito-borne viruses, such as dengue (DENV), chikungunya and zika virus, have developed into major public health challenges. Since all of these viruses may cause similar symptoms and can occur in concurrent epidemics, tools for their differential diagnosis and epidemiological monitoring are of urgent need. RESULTS: Here we report the application of a novel strategy to rapidly generate monoclonal antibodies (mAbs) against native viral antigens, exemplified for the DENV nonstructural glycoprotein 1 (NS1)...
November 22, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27858705/epitope-analysis-of-japanese-cedar-pollen-allergen-cry-j2-with-a-human-igm-class-monoclonal-antibody-404-117
#4
Seiya Chiba, Haruka Yokoyama, Ayane Kumazawa, Michie Shimmoto, Yasunori Naganawa, Hiroshi Shinmoto
Japanese cedar pollen allergen Cry j2 is a causal allergen of seasonal pollinosis in Japan. To analyze B cell epitopes of Cry j2, we established two human-mouse hybridomas secreting IgM class human monoclonal antibodies to Cry j2. A pin-peptide enzyme-linked immunosorbent assay with synthesized icosa peptides showed that 404-117 monoclonal antibody bound to peptides #11-13 with cry j2 amino acid sequence of 101F-L140. Detailed analysis with octa peptides and alanine substituted peptides indicated that an amino acid sequence of 118FKVD121 was an essential for antibody binding...
November 4, 2016: Human Antibodies
https://www.readbyqxmd.com/read/27845784/antibody-purification-via-affinity-membrane-chromatography-method-utilizing-nucleotide-binding-site-targeting-with-a-small-molecule
#5
Nur Mustafaoglu, Tanyel Kiziltepe, Basar Bilgicer
Here, we present an affinity membrane chromatography technique for purification of monoclonal and polyclonal antibodies from cell culture media of hybridomas and ascites fluids. The m-NBST method utilizes the nucleotide-binding site (NBS) that is located on the Fab variable domain of immunoglobulins to enable capturing of antibody molecules on a membrane affinity column via a small molecule, tryptamine, which has a moderate binding affinity to the NBS. Regenerated cellulose membrane was selected as a matrix due to multiple advantages over traditionally used resin-based affinity systems...
November 15, 2016: Analyst
https://www.readbyqxmd.com/read/27829125/development-of-a-new-anti-prolactin-receptor-prlr-antibody-f56-which-can-serve-as-a-prlr-antagonist
#6
Huanzhong Cui, Yun Zhi Ma, Yanrong Wang, Meng Song, Hui Zhang
In this work, we developed a new prolactin receptor (PRLR) antagonist using the hybridoma technique. A series of monoclonal antibodies against prolactin receptor (PRLR) were prepared, from which we characterized and selected one anti-PRLR antibody, F56. Epitome mapping showed that F56 and prolactin (PRL) share a common binding epitope on PRLR, and therefore, F56 could compete with prolactin (PRL) for binding to PRLR. Subsequent experiments indicated that F56 could effectively neutralize PRLR-mediated intracellular signalling molecules, such as signal transducer and activator of transcription (STAT) and extracellular signal-regulated kinase-1 and kinase 2 (ERK1/2), either by endogenously expressed PRLR or in a cell model transfected with PRLR...
November 6, 2016: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/27821304/a-novel-enzyme-linked-immunosorbent-assay-based-on-anti-lipovitellin-monoclonal-antibodies-for-quantification-of-zebrafish-danio-rerio-vitellogenin
#7
Jun Wang, Wei Wang, Hua Tian, Xiaona Zhang, Shaoguo Ru
Vitellogenin (Vtg) in zebrafish (Danio rerio) is a recommended biomarker endpoint for detecting estrogenic activity of chemicals under the OECD test guidelines. The present paper reports the development of a sensitive and rapid enzyme-linked immunosorbent assay (ELISA) for the quantification of zebrafish Vtg based on monoclonal antibodies (MAbs) against lipovitellin (Lv), the major yolk protein derived from Vtg. The purified Lv was used to immunize mice and the spleen cells of mice were fused with myeloma cells...
November 4, 2016: Ecotoxicology and Environmental Safety
https://www.readbyqxmd.com/read/27790631/hybridoma-cell-culture-and-glycan-profile-dataset-at-various-bioreactor-conditions
#8
Hemlata Bhatia, Erik Read, Cyrus Agarabi, Kurt Brorson, Scott Lute, Seongkyu Yoon
This is an "11 factor-2 level-12 run" Plackett-Burman experimental design dataset. The dataset includes 11 engineering bioreactor parameters as input variables. These 11 factors were varied at 2 levels and 23 response variables that are glycan profile attributes, were measured "A Design Space Exploration for Control of Critical Quality Attributes of mAb" (H. Bhatia, E.K. Read, C.D. Agarabi, K.A. Brorson, S.C. Lute, S. Yoon S, 2016) [2].
December 2016: Data in Brief
https://www.readbyqxmd.com/read/27788226/identification-of-immunoglobulin-gene-sequences-from-a-small-read-number-of-mrna-seq-using-hybridomas
#9
Yuki Kuniyoshi, Kazumitsu Maehara, Takeshi Iwasaki, Masayasu Hayashi, Yuichiro Semba, Masatoshi Fujita, Yuko Sato, Hiroshi Kimura, Akihito Harada, Yasuyuki Ohkawa
Identification of immunoglobulin genes in hybridomas is essential for producing antibodies for research and clinical applications. A couple of methods such as RACE and degenerative PCR have been developed for determination of the Igh and Igl/Igk coding sequences (CDSs) but it has been difficult to process a number of hybridomas both with accuracy and rapidness. Here, we propose a new strategy for antibody sequence determination by mRNA-seq of hybridomas. We demonstrated that hybridomas highly expressed the Igh and Igl/Igk genes and that de novo transcriptome assembly using mRNA-seq data enabled identification of the CDS of both Igh and Igl/Igk accurately...
2016: PloS One
https://www.readbyqxmd.com/read/27779884/de-novo-ms-ms-sequencing-of-native-human-antibodies
#10
Adrian Guthals, Yutian Gan, Laura Murray, Yongmei Chen, Jeremy Stinson, Gerald R Nakamura, Jennie R Lill, Wendy Sandoval, Nuno Bandeira
One direct route for the discovery of therapeutic human monoclonal antibodies (mAbs) involves the isolation of peripheral B cells from survivors/sero-positive individuals after exposure to an infectious reagent or disease etiology followed by single-cell sequencing or hybridoma generation. Peripheral B cells, however, are not always easy to obtain and only represent a small percentage of the total B cell population across all bodily tissues. Although it has been demonstrated that tandem mass spectrometry (MS/MS) techniques can interrogate the expressed polyclonal antibody (pAb) response to an antigen in vivo, all current approaches identify MS/MS spectra against databases derived from genetic sequencing of B cells from the same patient...
October 25, 2016: Journal of Proteome Research
https://www.readbyqxmd.com/read/27770198/production-and-characterization-of-a-monoclonal-antibody-specific-to-16%C3%A2-kda-antigen-of-paramphistomum-gracile
#11
Panat Anuracpreeda, Amaya Watthanadirek, Runglawan Chawengkirttikul, Prasert Sobhon
A number of monoclonal antibodies (MoAbs) against the 16 kDa antigen of Paramphistomum gracile (16 kDaAgPg) were produced in vitro by hybridoma technique. Reactivity and specificity of these MoAbs were evaluated by ELISA and immunoblotting assays. Seven MoAb clones were selected from the stable hybridoma clones, namely 1D10, 2D7, 3B10, 3D9, 4F1, 4G4, and 5G12. It was found to be IgM and kappa light chain isotypes. By immunoblotting and ELISA, all MoAbs reacted with purified 16 kDaAgPg at molecular weight (MW) of 16 kDa and with the native 16 kDa antigen at MW of 16 kDa in the whole body (WB) and excretory-secretory (ES) fractions, but not with tegumental antigens (TA) of adult fluke...
October 22, 2016: Parasitology Research
https://www.readbyqxmd.com/read/27738943/microencapsulated-cells-for-cancer-therapy
#12
L Saenz Del Burgo, J Ciriza, R M Hernández, G Orive, J L Pedraz
The microencapsulation of different types of cells that are able to produce therapeutic factors is being investigated for the treatment of several human diseases. Most efforts are focused on chronic and degenerative diseases as this strategy could become an alternative to some commonly used parenteral treatments that need to be repeatedly administered. But, this approach has also been investigated in the field of oncology with the aim of providing immunomodulatory antibodies that are able to enhance the patient's inherent immune response against the tumor...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27734371/preparation-of-module-specific-antibodies-against-ccn-family-members
#13
Satoshi Kubota, Masaharu Takigawa
Specific antibodies against biomolecules are conventional, but robust tools for the structural and functional analysis of target molecules. Since CCN family proteins are composed of four distinct modules that together determine the functionalities as full-length molecules depending upon extracellular microenvironment, specific antibody against independent modules are quite useful in CCN family research. Three distinct strategies are considerable for raising antibodies specific to four modules: IGFBP, VWC, TSP1, and CT modules...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27730560/optimized-generation-of-high-affinity-high-specificity-single-chain-fv-antibodies-from-multi-antigen-immunized-chickens
#14
William J J Finlay, Laird Bloom, Sreeja Varghese, Bénédicte Autin, Orla Cunningham
High-affinity, highly specific binding proteins are a key class of molecules used in the development of new affinity chromatography methods. Traditionally, antibody-based methods have relied on the use of immunoglobulins purified from immune animal sera, from egg yolks, or from murine monoclonal hybridoma supernatants. To accelerate and refine the reagent antibody generation process, we have developed optimized methods that allow the rapid assembly of scFv libraries from chickens immunized with pools of immunogens...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27706938/bispecific-monoclonal-antibody-based-multi-analyte-elisa-for-furaltadone-metabolite-malachite-green-and-leucomalachite-green-in-aquatic-products
#15
Feng Wang, Hong Wang, Yudong Shen, Yongjun Li, Jie-Xian Dong, Zhenlin Xu, Jinyi Yang, Yuanming Sun, Zhili Xiao
A new multi-analyte immunoassay was designed to screen furaltadone metabolite 5-morpholinomethyl-3-amino-2-oxazolidone (AMOZ), malachite green (MG), and leucomalachite green (LMG) in aquatic products using a bispecific monoclonal antibody (BsMAb). Gradient drug mutagenesis methods were separately used to prepare an anti-3-nitrobenzaldehyde-derivatized AMOZ (3-NPAMOZ) hybridoma cell line that was hypoxanthine-guanine-phosphoribosyltransferase (HGRPT) deficient and an anti-LMG hybridoma cell line that was thymidine kinase (TK) deficient...
October 5, 2016: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/27704358/a-humanized-chimeric-antibody-hai178-targeted-to-the-%C3%AE-subunit-of-f1f0-atp-synthase
#16
Chen Chen, Hui Liang, Xinmei Liao, Jian Pan, Jianhe Chen, Shibi Zhao, Yan Xu, Yun Wu, Jian Ni
Inhibition of tumor vasculature is an effective strategy for cancer therapy. Angiostatin could suppress tumor growth and metastasis by binding and inhibiting F1F0 ATP synthase on the endothelial cell surface. We previously screened a monoclonal antibody (McAb, McAb178-5G10), which specifically bound to ATPase on the surface of cells and showed an angiostatin-like activity. Here, we further generated a panel of CHO-mAb subclone stable expressing a humanized chimeric antibody from hybridoma cell McAb178-5G10 by gene engineer...
October 4, 2016: Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine
https://www.readbyqxmd.com/read/27689867/use-of-carbon-nanotubes-as-a-solid-support-to-establish-quantitative-centrifugation-and-qualitative-filtration-immunoassays-to-detect-gentamicin-contamination-in-commercial-milk
#17
Kun Zeng, Wei Wei, Ling Jiang, Fang Zhu, Daolin Du
Current methods to detect gentamicin (GEN), a broad-spectrum antibiotic that causes ototoxicity and nephrotoxicity when present in excess, have several limitations. Hence, we have developed two methods using multi-walled carbon nanotubes (MWCNTs) as a solid support to detect GEN. Hybridoma cells (2D12) producing high-sensitivity antibodies against GEN were established. The goat anti-mouse antibody was immobilized on MWCNTs directly or using bifunctional polyethylene glycol as a linker. On the basis of the physical characteristics of MWCNTs, a quantitative method involving centrifugation separation and a qualitative method involving filtration separation were established...
October 11, 2016: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/27678288/expression-and-purification-of-mouse-ttyh1-fragments-as-antigens-to-generate-ttyh1-specific-monoclonal-antibodies
#18
Xiu-Li Cao, Xing Zhang, Yu-Fei Zhang, Yi-Zhe Zhang, Chang-Geng Song, Fan Liu, Yi-Yang Hu, Min-Hua Zheng, Hua Han
Ttyh1 is a murine homolog of the Drosophila Tweety and is predicted as a five-pass transmembrane protein. The Ttyh1 mRNA is expressed in mouse brain tissues with a restricted pattern and in human glioma cells. Ttyh1 protein may function as a large-conductance chloride channel, however, the role of Ttyh1 in normal neural development and tumorigenesis has been largely unknown, at least partially due to the lack of effective antibodies. Here we report the expression in E. coli and purification of two recombinant Ttyh1 protein fragments corresponding to one of the predicted extracellular domains and the carboxyl terminus of the mouse Ttyh1...
September 25, 2016: Protein Expression and Purification
https://www.readbyqxmd.com/read/27677608/a-novel-fusion-protein-achr-fc-ameliorates-myasthenia-gravis-by-neutralizing-antiacetylcholine-receptor-antibodies-and-suppressing-acetylcholine-receptor-reactive-b-cells
#19
Masayuki Homma, Akiyuki Uzawa, Hitoshi Tanaka, Naoki Kawaguchi, Tetsuya Kanai, Kenji Nakajima, Masakuni Narita, Yukio Hara, Hideya Maruyama, Yasumasa Ogawa, Keiichi Himuro, Satoshi Kuwabara
Most patients with myasthenia gravis (MG) have elevated levels of autoantibodies against the nicotinic acetylcholine receptor (AChR) at the neuromuscular junction, which leads to muscle weakness. We developed a fusion protein, AChR-Fc, as a novel therapeutic biomolecule for patients with MG and examined its efficacy. AChR-Fc was expressed by Chinese hamster ovary cells and purified. We examined the neutralizing activity and cellular cytotoxicity of AChR-Fc using anti-AChR antibody-producing hybridoma cells and serum samples from 16 patients with MG...
September 27, 2016: Neurotherapeutics: the Journal of the American Society for Experimental NeuroTherapeutics
https://www.readbyqxmd.com/read/27669301/glypican-3-targeting-immunotoxins-for-the-treatment-of-liver-cancer
#20
REVIEW
Bryan D Fleming, Mitchell Ho
Hepatocellular carcinoma (HCC) is the most common form of primary liver cancer, yet no effective therapeutics exist. This review provides an overview of the recent development of recombinant immunotoxins for the treatment of glypican-3 (GPC3) expressing HCC. GPC3 is a cell surface heparan sulfate proteoglycan that is overexpressed in HCC, but is absent from normal adult human tissues. Treatment of HCC with anti-GPC3 immunotoxins represents a new therapeutic option. Using phage display and hybridoma technologies, three high affinity antibodies (HN3, HS20 and YP7) have been generated against GPC3...
2016: Toxins
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