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https://www.readbyqxmd.com/read/28186433/production-of-monoclonal-antibody-against-excretory-secretory-antigen-of-fasciola-hepatica-and-evaluation-of-its-efficacy-in-the-diagnosis-of-fascioliasis
#1
Samaneh Abdolahi Khabisi, Bahador Sarkari, Abdolali Moshfe, Sedigheh Jalali
Parasitological methods are not helpful for the diagnosis of fascioliasis in acute and invasive periods of the disease. Detection of coproantigens seems to be a suitable alternative approach in the diagnosis of fascioliasis. The present study aimed to develop a reliable antigen detection system, using monoclonal antibodies raised against excretory-secretory (ES) antigen of Fasciola hepatica, for the diagnosis of fascioliasis. Fasciola adult worms were collected from the bile ducts of infected animals. Species of the fluke was determined by polymerase chain reaction-restriction fragment length polymorphism (RFLP-PCR)...
February 10, 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28154257/antibody-fragments-for-on-site-testing-of-cannabinoids-generated-via-in-vitro-affinity-maturation
#2
Izumi Morita, Hiroyuki Oyama, Mayumi Yasuo, Kazuhisa Matsuda, Kengo Katagi, Aya Ito, Hiroka Tatsuda, Hiroyuki Tanaka, Satoshi Morimoto, Norihiro Kobayashi
Law enforcement against illicit use of cannabis and related substances requires rapid, feasible, and reliable tools for on-site testing of cannabinoids. Notably, methods based on cannabinoid-specific antibodies enable efficient screening of multiple specimens. Antibody engineering may accelerate development of modern and robust testing systems. Here, we used in vitro affinity maturation to generate a single-chain Fv fragment (scFv) that recognizes with high affinity the psychoactive cannabinoid, Δ(9)-tetrahydrocannabinol (THC)...
2017: Biological & Pharmaceutical Bulletin
https://www.readbyqxmd.com/read/28144646/an-open-sandwich-immunoassay-for-detection-of-13-r-s-hydroxy-9-e-11-e-octadecadienoic-acid
#3
Jinhua Dong, Mototada Shichiri, Chan-I Chung, Takahiro Shibata, Koji Uchida, Yoshihisa Hagihara, Yasukazu Yoshida, Hiroshi Ueda
Lipid peroxidation is involved in many disorders and diseases such as cardiovascular disease, cancers, neurodegenerative diseases, and even aging. Lipid peroxidation products existing in blood or bodily fluids are very important biomarkers for the diagnosis of such diseases. In particular, 13(R,S)-hydroxy-9(E),11(E)-octadecadienoic acid (13-(E,E)-HODE) is an oxidiation product of linoleic acid, which is an important biomarker for many diseases such as diabetes and Alzheimer's disease. In this study, we successfully displayed the antigen-binding fragment of an antibody produced by hybridoma 1213-1 on the M13 phage and performed analysis of the antibody variable region genes...
February 1, 2017: Analyst
https://www.readbyqxmd.com/read/28137617/bioprocess-optimization-under-uncertainty-using-ensemble-modeling
#4
Yang Liu, Rudiyanto Gunawan
The performance of model-based bioprocess optimizations depends on the accuracy of the mathematical model. However, models of bioprocesses often have large uncertainty due to the lack of model identifiability. In the presence of such uncertainty, process optimizations that rely on the predictions of a single "best fit" model, e.g. the model resulting from a maximum likelihood parameter estimation using the available process data, may perform poorly in real life. In this study, we employed ensemble modeling to account for model uncertainty in bioprocess optimization...
January 27, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28125314/bisfabs-tools-for-rapidly-screening-hybridoma-iggs-for-their-activities-as-bispecific-antibodies
#5
Sanket Patke, Ji Li, Peiyin Wang, Dion Slaga, Jennifer Johnston, Sunil Bhakta, Siler Panowski, Liping L Sun, Teemu Junttila, Justin M Scheer, Diego A Ellerman
Bispecific antibodies are a growing class of therapeutic molecules. Many of the current bispecific formats require DNA engineering to convert the parental monoclonal antibodies into the final bispecific molecules. We describe here a method to generate bispecific molecules from hybridoma IgGs in 3-4 days using chemical conjugation of antigen-binding fragments (Fabs) (bisFabs). Proteolytic digestion conditions for each IgG isotype were analyzed to optimize the yield and quality of the final conjugates. The resulting bisFabs showed no significant amounts of homodimers or aggregates...
January 26, 2017: MAbs
https://www.readbyqxmd.com/read/28122974/monoclonal-antibodies-to-v2-v3-the-cd4-binding-site-and-gp41-hiv-1-mediate-phagocytosis-in-a-dose-dependent-manner
#6
Thomas Musich, Liuzhe Li, Lily Liu, Susan Zolla-Pazner, Marjorie Robert-Guroff, Miroslaw K Gorny
: In light of weak or absent neutralizing activity mediated by anti-V2 monoclonal Abs (mAbs), we tested whether they can mediate Ab-dependent cellular phagocytosis (ADCP) which is an important element of anti-HIV-1 immunity. We tested six anti-V2 mAbs and compared them with 21 mAbs specific for V3, the CD4-binding site (CD4bs), and gp41 derived from HIV-1 chronically infected individuals and produced by hybridoma cells. ADCP activity was measured by flow cytometry using uptake by THP-1 monocytic cells of fluorescent beads coated with gp120, gp41, BG505 SOSIP...
January 25, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28119207/epitope-dependent-mechanisms-of-cd27-neutralization-revealed-by-x-ray-crystallography
#7
Galina Obmolova, Alexey Teplyakov, Thomas J Malia, Nicole Wunderler, Deborah Kwok, Linda Barone, Raymond Sweet, Tatiana Ort, Michael Scully, Gary L Gilliland
CD27 is a T and B cell co-stimulatory protein of the TNF receptor superfamily dependent on the availability of the TNF-like ligand CD70. Two anti-CD27 neutralizing monoclonal antibodies were obtained from mouse hybridoma and subsequently humanized and optimized for binding the target. The two antibodies are similar in terms of their CD27-binding affinity and ability to block NF-κB signaling, however their clearance rates in monkeys are very different. The pharmacokinetics profiles could be epitope dependent...
March 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28117810/generation-of-murine-monoclonal-antibodies-by-hybridoma-technology
#8
Pamela Holzlöhner, Katja Hanack
Monoclonal antibodies are universal binding molecules and are widely used in biomedicine and research. Nevertheless, the generation of these binding molecules is time-consuming and laborious due to the complicated handling and lack of alternatives. The aim of this protocol is to provide one standard method for the generation of monoclonal antibodies using hybridoma technology. This technology combines two steps. Step 1 is an appropriate immunization of the animal and step 2 is the fusion of B lymphocytes with immortal myeloma cells in order to generate hybrids possessing both parental functions, such as the production of antibody molecules and immortality...
January 2, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28115917/a-new-enzyme-linked-immunosorbent-assay-elisa-for-human-free-and-bound-kallikrein-9
#9
Panagiota Filippou, Dimitrios Korbakis, Sofia Farkona, Antoninus Soosaipillai, Theano Karakosta, Eleftherios P Diamandis
BACKGROUND: Kallikrein 9 (KLK9) is a member of the human kallikrein-related peptidases family, whose physiological role and implications in disease processes remain unclear. The active form of the enzyme is predicted to have chymotryptic activity. In the present study, we produced for the first time the active recombinant protein and monoclonal antibodies, and developed novel immunoassays for the quantification of free and bound KLK9 in biological samples. METHODS: The coding sequence of mature KLK9 isoform (mat-KLK9) was expressed in an Expi293F mammalian system and the synthesized polypeptide was purified through a two-step protocol...
2017: Clinical Proteomics
https://www.readbyqxmd.com/read/28115505/identification-of-mycobacterial-rplj-l10-and-rpsa-s1-proteins-as-novel-targets-for-cd4-t-cells
#10
Alison J Johnson, Steven C Kennedy, Cecilia S Lindestam Arlehamn, Michael F Goldberg, Neeraj K Saini, Jiayong Xu, Sinu Paul, Subray S Hegde, John S Blanchard, John Chan, William R Jacobs, Alessandro Sette, Steven A Porcelli
Tuberculosis (TB) due to Mycobacterium tuberculosis (Mtb) remains a major global infectious disease problem, and a more efficacious vaccine is urgently needed for control and prevention of disease caused by this organism. We previously reported a genetically modified strain of Mycobacterium smegmatis called IKEPLUS as a promising TB vaccine candidate. Since protective immunity induced by IKEPLUS is dependent on antigen-specific CD4(+) T cell memory, we hypothesized that the specificity of the CD4(+) T cell response was a critical feature of this protection...
January 23, 2017: Infection and Immunity
https://www.readbyqxmd.com/read/28109156/-cloning-and-sequence-analysis-of-variable-region-genes-of-mouse-anti-human-s100a9-monoclonal-antibody
#11
Min Yan, Danyang Mi, Xiangdong Sun, Zhenqiang Zhang, Huahui Zeng, Wendi Liu
Objective To clone and analyze variable region gene sequences of mouse monoclonal antibody against human S100A9 (anti-human S100A9 mAb). Methods Total RNA was extracted from the hybridoma cell line (IID4B10) secreting mouse anti-human S100A9 mAb. Furthermore, the heavy chain variable region (VH) and light chain variable region (VL) genes were amplified using reverse transcription-polymerase chain reaction (RT-PCR). After the cloning and sequencing of the VH and VL genes, the sequence analyses were performed by BLAST program...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28107434/anti-mrka-monoclonal-antibodies-reveal-distinct-structural-and-antigenic-features-of-mrka
#12
Qun Wang, Yan Chen, Romana Cvitkovic, Meghan E Pennini, Chew Shun Chang, Mark Pelletier, Jessica Bonnell, Adem C Koksal, Herren Wu, William F Dall'Acqua, C Kendall Stover, Xiaodong Xiao
Antibody therapy against antibiotics resistant Klebsiella pneumoniae infections represents a promising strategy, the success of which depends critically on the ability to identify appropriate antibody targets. Using a target-agnostic strategy, we recently discovered MrkA as a potential antibody target and vaccine antigen. Interestingly, the anti-MrkA monoclonal antibodies isolated through phage display and hybridoma platforms all recognize an overlapping epitope, which opens up important questions including whether monoclonal antibodies targeting different MrkA epitopes can be generated and if they possess different protective profiles...
2017: PloS One
https://www.readbyqxmd.com/read/28100678/modulation-of-naive-cd8-t-cell-response-features-by-ligand-density-affinity-and-continued-signaling-via-internalized-tcrs
#13
Renu Balyan, Rupali Gund, Chitra Ebenezer, Jasneet Kaur Khalsa, Divya Anna Verghese, Thyagarajan Krishnamurthy, Anna George, Vineeta Bal, Satyajit Rath, Ashutosh Chaudhry
T cell response magnitudes increase with increasing antigenic dosage. However, it is unclear whether ligand density only modulates the proportions of responding ligand-specific T cells or also alters responses at the single cell level. Using brief (3 h) exposure of TCR-transgenic mouse CD8 T cells in vitro to varying densities of cognate peptide-MHC ligand followed by ligand-free culture in IL-2, we found that ligand density determined the frequencies of responding cells but not the expression levels of the early activation marker molecule, CD69...
January 18, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28092005/investigation-of-diethylstilbestrol-residue-level-in-human-urine-samples-by-a-specific-monoclonal-antibody
#14
Dai Zhang, Lifang Zhou, Yajing Lei, Zhan Zhou, Jie Zhou, Shuqing Chen
Diethylstilbestrol (DES) is used as a kind of animal feed additive and affects people's health through the food chain. The purpose of this study is to detect the residue level of DES in 576 human urine samples directly. DES-BSA was used to immunize Balb/c mice. The monoclonal antibody was produced by hybridoma that was screened through cell fusion techniques. Finally, we developed the indirect competitive ELISA method to analyze 576 human urine samples from Zhejiang Province, China. The IC50 of this method was 3...
January 14, 2017: Environmental Science and Pollution Research International
https://www.readbyqxmd.com/read/28043495/development-of-rapid-immuno-diagnostic-test-for-the-early-detection-of-tuberculosis
#15
Anil Kumar Gupta, Saumya Srivastava, Amit Singh, Sarman Singh
AIMS AND OBJECTIVE: To develop novel immunodiagnostic test for tuberculosis. METHODS: The selected novel proteins named as Rv2145c (SS1), Rv1437 (SS2), Rv1827 (SS3), and Rv2970c (SS4) were cloned, expressed and purified under specific conditions. Additionally, monoclonal antibodies (mAbs) were developed using these recombinant antigens via hybridoma technology. Hybridoma clones were screened and positive clones were selected for further experiment. The mAbs were purified from cell culture supernatant using protein A/G column chromatography...
December 2016: International Journal of Mycobacteriology
https://www.readbyqxmd.com/read/28031526/radiolabeled-novel-mab-4g1-for-immunospect-imaging-of-egfrviii-expression-in-preclinical-glioblastoma-xenografts
#16
Xujie Liu, Chengyan Dong, Jiyun Shi, Teng Ma, Zhongxia Jin, Bing Jia, Zhaofei Liu, Li Shen, Fan Wang
Epidermal growth factor receptor mutant III (EGFRvIII) is exclusively expressed in tumors, such as glioblastoma, breast cancer and hepatocellular carcinoma, but never in normal organs. Increasing evidence suggests that EGFRvIII has clinical significance in glioblastoma prognosis due to its enhanced tumorigenicity and chemo/radio resistance, thus the development of an imaging approach to early detect EGFRvIII expression with high specificity is urgently needed. To illustrate this point, we developed a novel anti-EGFRvIII monoclonal antibody 4G1 through mouse immunization, cell fusion and hybridoma screening and then confirmed its specificity and affinity by a serial of assays...
January 24, 2017: Oncotarget
https://www.readbyqxmd.com/read/28029330/preparation-and-identification-of-monoclonal-antibody-against-c1q-tnf-related-protein-4
#17
He Huang, Xiaotong Wu, Lulu Cao, Lu Wang
CTRP4 (C1q/TNF-related protein 4) is a secreted cytokine homologous to adiponectin, which plays an important role in immunity and metabolism. This study was designed to generate CTRP4 monoclonal antibodies (MAbs). Splenocytes extracted from mice immunized with prokaryotic protein were fused with myeloma cells Sp2/0 to generate hybridoma cells. Three hybridoma strains (16, 33, and 35) were chosen and their MAbs were purified. The specificity and affinity were identified by Western blot and enzyme-linked immunosorbent assay, whereas CTRP4 endogenous expression was identified by immunofluorescence staining...
December 2016: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28027008/development-of-an-immunoassay-using-an-anti-wogonin-glucuronide-monoclonal-antibody
#18
Madan Kumar Paudel, Seiichi Sakamoto, Le Van Huy, Hiroyuki Tanaka, Miyamoto Tomofumi, Takano Akihito, Satoshi Morimoto
Wogonin 7-O-β-D-glucuronide (Wgn) is a bioactive flavone present in the dried root of Scutellaria baicalensis Georgi. To generate a monoclonal antibody (MAb) against Wgn, BALB/c mice injected with Wgn-bovine serum albumin yielded splenocytes that we fused with SP2/0 myeloma cells using the polyethylene glycol method. We obtained a hybridoma designated 315A that produced a MAb reactive to Wgn. The anti-Wgn MAb 315A was applied to an indirect competitive enzyme-linked immunosorbent assay (icELISA) to quantify Wgn...
December 27, 2016: Journal of Immunoassay & Immunochemistry
https://www.readbyqxmd.com/read/28026995/scale-up-production-and-characterization-of-anti-human-cardiac-troponin-i-monoclonal-antibody-in-ascitic-fluid-of-balb-c-mice
#19
Asiabanha Rezaee Majid, Rasaee Mohammad Javad, Paknejad Malihe, Mohammadnejad Javad
The Human Cardiac Troponin I (hcTnI) is a 210 amino acid protein, 23 kDa in molecular weight. This biomarker is commonly used to diagnose myocardial infarction, micro injury, and acute coronary syndrome (ACS) in patients referring to emergency departments. The American Heart Association (AHA) and European Society of Cardiology (ESC) proposed troponin I as the gold biomarker for early detection of heart attack, especially in myocardial infarction (MI). Therefore, developing monoclonal antibodies against this biomarker could help in for early detection of heart attack...
December 27, 2016: Journal of Immunoassay & Immunochemistry
https://www.readbyqxmd.com/read/28013222/generation-and-characterization-of-monoclonal-antibodies-against-human-lgr6
#20
Shin-Ichi Funahashi, Yasunori Suzuki, Kiyotaka Nakano, Shigeto Kawai, Masami Suzuki
Leucine-rich repeat-containing G protein-coupled receptor 6 (LGR6) is a seven-pass transmembrane protein known to be a marker of stem cells in several organs. To deepen our understanding of the cell biology of LGR6-positive cells, including stem cells, we generated monoclonal antibodies (mAbs) against human LGR6. DNA immunization followed by whole-cell immunization with LGR6-expressing transfectants was performed to obtain mAbs that recognized the native form of LGR6. Hybridomas were screened by flow cytometry using LGR6-transfected cells...
December 24, 2016: Journal of Biochemistry
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