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Journal of Immunological Methods

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https://www.readbyqxmd.com/read/28931470/free-light-chains-eclectic-multipurpose-biomarker
#1
REVIEW
Umberto Basile, Francesca Gulli, Laura Gragnani, Cecilia Napodano, Krizia Pocino, Gian Ludovico Rapaccini, Michele Mussap, Anna Linda Zignego
The production of antibodies is accompanied by a slight excess of synthesis of κ and λ immunoglobulin light chains; small amounts of them are released in the peripheral blood and can also be found in various body fluids, such as synovial fluid, cerebrospinal fluid, urine and saliva. They are rapidly filtered by the glomerulus and >99% are reabsorbed from the cells of the proximal convoluted tubule, making them present in the urine in only trace amounts. The production of an excess of protein without a reason or a specific function in a biological system is rare...
September 17, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28927728/cryopreservation-of-human-whole-blood-allows-immunophenotyping-by-flow-cytometry-up-to-30days-after-cell-isolation
#2
R Madelaine Paredes, Douglas K Tadaki, Amanda Sooter, Fabia Gamboni, C D R Forest Sheppard
Immunophenotyping of whole blood (WB) by flow cytometry (FC) is used clinically to assess a patient's immune status and also in biomedical research. Current protocols recommend storage of immunolabeled samples at 4°C with FC analysis to be completed within seven days. This data acquisition window can be extended to up to one year post-labeling, but this requires cryopreservation of the samples at ultra-low temperatures (≤-80°C or in liquid nitrogen). In this study we optimized a standardized cryopreservation protocol to enable preservation of immunolabeled, human WB samples at -20°C for FC and tested its effectiveness after 0, 5, 15 or 30days...
September 16, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28919468/a-copper-based-enzyme-free-fluorescence-elisa-for-her2-detection
#3
Shiyu Tian, Ke Zeng, Aijun Yang, Qin Wang, Minghui Yang
We reported an enzyme-free ELISA to detect breast cancer biomarker human epidermal growth factor receptor 2 (HER2) in human serum samples. Instead of enzymes (such as horseradish peroxidase) used in traditional ELISA, CuO nanoparticles were utilized as the signal probe. Compared to traditional enzymes, CuO nanoparticles have the advantages of low cost and good stability. After dissolving CuO nanoparticles with acid, the Cu (II) ions generated catalyzed the reaction of o-phenylenediamine with ascorbic acid to produce fluorescent quinoxaline derivative molecules...
September 14, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28917434/quantitative-analysis-of-four-protein-biomarkers-an-automated-microfluidic-cartridge-based-method-and-its-comparison-to-colorimetric-elisa
#4
REVIEW
Mark Dysinger, Greg Marusov, Stephanie Fraser
Biomarker quantitation with ligand binding assays has matured greatly in recent years. This maturation has been partly in response to demands for more data points from fewer samples or less available sample volume. Multiplexing offers opportunities to acquire data for multiple analytes from single sample assay iterations, but has its own unique challenges and limitations. ProteinSimple has developed Simple Plex™, an automated immunoassay platform consisting of microfluidic cartridge-based assays run on the Ella instrument...
September 13, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28890365/detection-and-quantification-of-rituximab-in-the-human-urine
#5
Roland Jacobs, Thais Langer-Jacobus, Michelle Duong, Klaus Stahl, Hermann Haller, Reinhold E Schmidt, Mario Schiffer
B cell depletion by rituximab treatment might be inefficient in patients suffering from nephrotic syndrome. Due to the impaired glomerular filtration barrier a significant portion of the therapeutic antibody might be lost into the urinary space. In order to determine the amount of rituximab in the urine of such patients, CD20+ Daudi cells were stained with the patients' urine followed by a fluorochrome-labeled secondary antibody. Mean fluorescence intensity of that way labeled Daudi cells was determined by flow cytometry...
September 7, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28890364/sequential-screening-by-clonepix-fl-and-intracellular-staining-facilitate-isolation-of-high-producer-cell-lines-for-monoclonal-antibody-manufacturing
#6
Gargi Roy, Guillermo Miro-Quesada, Li Zhuang, Tom Martin, Jie Zhu, Herren Wu, Marcello Marelli, Michael A Bowen
The generation of stable cell lines that express therapeutic monoclonal antibodies (mAbs) for manufacturing purposes is a time and labor intensive process. Since cell line development is on the critical path to clinical testing, timely generation of highly productive manufacturing cell lines is essential to remain competitive in the biopharmaceutical industry. To aid this essential function we have developed a methodology for the selection of antibody expressing cells using fluorescence based ClonePix FL colony isolation and flow cytometry analysis following intracellular staining for immunoglobulin G (IgG)...
September 7, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28882613/average-overlap-frequency-a-simple-metric-to-evaluate-staining-quality-and-community-identification-in-high-dimensional-mass-cytometry-experiments
#7
El-Ad David Amir, Xinzheng V Guo, Oksana Mayovska, Adeeb Rahman
High dimensional cytometry now allows measurement of over 50 parameters in a single sample, and is typically visualized using sophisticated dimensionality-reducing methods and analyzed with automated clustering algorithms. While these tools facilitate the identification and presentation of key findings, it remains challenging to effectively monitor and report the staining quality of individual markers. We present the Average Overlap Frequency (AOF), a simple and efficient metric to evaluate and quantify the robustness of staining and clustering quality in high-dimensional data...
September 4, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28882612/proposed-panel-of-diagnostic-tools-for-accurate-temporal-classification-of-symptomatic-t-gondii-infection
#8
Geisa Baptista Barros, Elenice Moreira Lemos, Priscila Pinto E Silva-Dos-Santos, Reynaldo Dietze, Eliana Zandonade, José Roberto Mineo, Deise Aparecida de Oliveira Silva, Ana Cláudia Marquez Pajuaba, Matheus de Souza Gomes, Laurence Rodrigues do Amaral, Jordana Grazziela Coelho-Dos-Reis, Olindo Assis Martins-Filho, José Carlos Serufo
Serological tests available for the diagnosis of acute Toxoplasma gondii infection have limitations in establishing the temporal diagnosis of acute toxoplasmosis. The present analytical-descriptive investigation comprises of a prospective longitudinal cohort study to search for accurate biomarkers to distinguish acute, early and late convalescent T. gondii infection. Classic methods (immunofluorescence-IFA along with Enzyme-linked immunosorbent-ELISA and fluorescent-ELFA assays) for IgM, IgA, IgG and IgG avidity were employed in parallel with flow cytometry-based anti-fixed T...
September 4, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28882611/generation-and-crispr-cas9-editing-of-transformed-progenitor-b-cells-as-a-pseudo-physiological-system-to-study-dna-repair-gene-function-in-v-d-j-recombination
#9
Hélène Lenden Hasse, Chloé Lescale, Joy J Bianchi, Wei Yu, Marie Bedora-Faure, Ludovic Deriano
Antigen receptor gene assembly is accomplished in developing lymphocytes by the V(D)J recombination reaction, which can be separated into two steps: DNA cleavage by the recombination-activating gene (RAG) nuclease and joining of DNA double strand breaks (DSBs) by components of the nonhomologous end joining (NHEJ) pathway. Deficiencies for NHEJ factors can result in immunodeficiency and a propensity to accumulate genomic instability, thus highlighting the importance of identifying all players in this process and deciphering their functions...
September 4, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28855106/sample-processing-approach-for-detection-of-ricin-in-surface-samples
#10
Staci Kane, Sanjiv Shah, Anne Marie Erler, Teneile Alfaro
With several ricin contamination incidents reported over the past decade, rapid and accurate methods are needed for environmental sample analysis, especially after decontamination. A sample processing method was developed for common surface sampling devices to improve the limit of detection and avoid false negative/positive results for ricin analysis. Potential assay interferents from the sample matrix (bleach residue, sample material, wetting buffer), including reference dust, were tested using a Time-Resolved Fluorescence (TRF) immunoassay...
August 27, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28855105/multiplex-flow-cytometry-based-assay-to-study-the-breadth-of-antibody-responses-against-e1e2-glycoproteins-of-hepatitis-c-virus
#11
Sabrina J Merat, Dorien van de Berg, Camille Bru, Etsuko Yasuda, Esther Breij, Neeltje Koostra, Maria Prins, Richard Molenkamp, Arjen Q Bakker, Menno D de Jong, Hergen Spits, Janke Schinkel, Tim Beaumont
Hepatitis C virus (HCV) infection is a major global public health problem. In infected subjects who clear HCV infection, a strong antibody response has proven to be a correlate of protection. Understanding the antibody response in multiple subjects in large-scale studies would greatly benefit vaccine development. To determine the breadth of a polyclonal-serum antibody response, and or, the monoclonal antibodies against the different HCV E1E2 genotypes, we developed a quick and high throughput flow cytometry assay using fluorescent cell barcoding to distinguish cells transfected with different E1E2 sequences in a single measurement...
August 27, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28847736/development-of-a-luciferase-reporter-jurkat-cell-line-under-the-control-of-endogenous-interleukin-2-promoter
#12
Jinqi Liu, Ren Liu, Peter Gray, Zhenyi Liu, Xiaoxia Cui, Guanghua Li, Zhong Liu
During new drug development, it is critical to have a cell-based reporter bioassay to measure drug-mediated physiological changes. In a conventional reporter cell line, a reporter expression construct is randomly inserted into the host cell genome with the reporter gene under control of an engineered promoter. This design ensures high signal output but may not represent the true physiological cell signaling. Here we used the CRISPR/Cas9 technology to engineer a Jurkat cell line by replacing one interleukin 2 (IL2) allele with firefly luciferase gene while keeping the other IL2 allele intact...
August 26, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28827190/establishing-tools-for-early-diagnosis-of-congenital-toxoplasmosis-flow-cytometric-igg-avidity-assay-as-a-confirmatory-test-for-neonatal-screening
#13
Aline de Castro Zacche-Tonini, Giuliana Schmidt França Fonseca, Laura Néspoli Nassar Passini de Jesus, Geisa Baptista Barros, Jordana Grazziela Alves Coelho-Dos-Reis, Samantha Ribeiro Béla, Anderson Silva Machado, Ana Carolina Aguiar Vasconcelos Carneiro, Gláucia Manzan Queiroz Andrade, Daniel Vitor Vasconcelos-Santos, José Nélio Januário, Andréa Teixeira-Carvalho, Ricardo Wagner Almeida Vitor, Eloísa Amália Vieira Ferro, José Roberto Mineo, Olindo Assis Martins-Filho, Elenice Moreira Lemos
The aim of this study was to evaluate the performance of conventional serology (Q-Preven™ and ELFAVIDAS™) and flow cytometry-based serologic tools for early serologic diagnosis of congenital toxoplasmosis. The study groups included prospectively confirmed cases of congenital toxoplasmosis (TOXO=88) and age-matching non-infected controls (NI=15).The results demonstrated that all samples tested positive/indeterminate for anti-T. gondii IgM screening at birth using air-dried whole blood samples. Serum samples collected at 30-45days after birth tested positive for ELFAVIDAS™ IgG in both groups...
August 18, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28827189/prediction-of-antibody-structural-epitopes-via-random-peptide-library-screening-and-next-generation-sequencing
#14
Kelly N Ibsen, Patrick S Daugherty
Next generation sequencing (NGS) is widely applied in immunological research, but has yet to become common in antibody epitope mapping. A method utilizing a 12-mer random peptide library expressed in bacteria coupled with magnetic-based cell sorting and NGS correctly identified >75% of epitope residues on the antigens of two monoclonal antibodies (trastuzumab and bevacizumab). PepSurf, a web-based computational method designed for structural epitope mapping was utilized to compare peptides in libraries enriched for monoclonal antibody (mAb) binders to antigen surfaces (HER2 and VEGF-A)...
August 18, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28803843/high-throughput-screening-of-hybridoma-supernatants-using-multiplexed-fluorescent-cell-barcoding-on-live-cells
#15
Mei Lu, Brian M Chan, Peter W Schow, Wesley S Chang, Chadwick T King
With current available assay formats using either immobilized protein (ELISA, enzyme-linked immunosorbent assay) or immunostaining of fixed cells for primary monoclonal antibody (mAb) screening, researchers often fail to identify and characterize antibodies that recognize the native conformation of cell-surface antigens. Therefore, screening using live cells has become an integral and important step contributing to the successful identification of therapeutic antibody candidates. Thus the need for developing high-throughput screening (HTS) technologies using live cells has become a major priority for therapeutic mAb discovery and development...
August 10, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28802832/retroviral-gene-transfer-into-primary-human-nk-cells-activated-by-il-2-and-k562-feeder-cells-expressing-membrane-bound-il-21
#16
Maria A Streltsova, Eugene Barsov, Sofia A Erokhina, Elena I Kovalenko
Natural killer (NK) cells are capable of rapidly recognizing and efficiently killing tumor cells. This makes them a potentially promising agent for cancer immunotherapy. Additional genetic modifications of NK cells may further improve their anti-tumor efficacy. Numerous technical challenges associated with gene delivery into NK cells have significantly tempered this approach. We achieved efficient retroviral vector transduction of primary human NK cells that were stimulated by a combination of IL-2 and engineered K562 cells expressing membrane-bound IL-21...
August 10, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28789924/constitutive-interaction-between-4-1bb-and-4-1bbl-on-murine-lps-activated-bone-marrow-dendritic-cells-masks-detection-of-4-1bbl-by-tks-1-but-not-19h3-antibody
#17
Achire N Mbanwi, Gloria H Y Lin, Kuan Chung Wang, Tania H Watts
4-1BB is a TNFR family member associated with NF-κB mediated survival signaling. 4-1BB is widely expressed on activated cells of the immune system, including activated T cells, NK cells and dendritic cells. Its ligand, 4-1BBL, is transiently expressed on activated antigen presenting cells and at low levels on activated T cells. Although 4-1BBL-deficient mice clearly demonstrate a role for 4-1BBL in CD8 T cell responses to viruses such as influenza, 4-1BBL can be difficult to detect following infection of mice...
August 5, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28782523/monitoring-native-hla-i-trimer-specific-antibodies-in-luminex-multiplex-single-antigen-bead-assay-evaluation-of-beadsets-from-different-manufacturers
#18
Mepur H Ravindranath, Vadim Jucaud, Soldano Ferrone
Luminex single antigen bead (SAB) assay utilizes beadsets coated with a set of cloned and purified HLA molecules, for monitoring serum anti-HLA antibodies. Particularly, the level of serum IgG against native HLA-I trimers (heavy chain (HC) and β2-microglobulin (β2m) with a peptide), expressed in allograft tissues is correlated with graft failure. In addition to native trimeric HLAI, the beadsets may carry HC only or the dimeric variants, peptide-free HC with β2m and β2m-free HC with or without peptides...
August 4, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28760669/an-immunoproteomic-approach-revealed-antigenic-proteins-enhancing-serodiagnosis-performance-of-bird-fancier-s-lung
#19
Adeline Rouzet, Gabriel Reboux, Jean-Charles Dalphin, Anne Gondouin, Paul De Vuyst, Thierry Balliau, Laurence Millon, Benoit Valot, Sandrine Roussel
BACKGROUND: Bird fancier's lung (BFL) caused by repeated inhalation of avian proteins is the most common form of hypersensitivity pneumonitis. However, the exact identification of proteins involved is unknown, and serological test use for diagnosis need to be standardized. The objectives of this study were (i) to identify antigenic proteins from pigeon droppings (ii) to provide information about their location in avian matrices and (iii) to produce them in recombinant proteins to evaluate their diagnostic performances...
July 29, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28760671/application-of-phospho-cytof-to-characterize-immune-activation-in-patients-with-sickle-cell-disease-in-an-ex-vivo-model-of-thrombosis
#20
Jeffrey Glassberg, Adeeb H Rahman, Mohammad Zafar, Caroline Cromwell, Alexa Punzalan, Juan Jose Badimon, Louis Aledort
Sickle cell disease (SCD) is a genetic disease caused by mutations in the beta globin gene, and inflammation plays a key role in driving many aspects of disease pathology. Early immune activation is believed to be associated with hemodynamic stresses and thrombus formation as cells traffic through blood vessels. We applied an extracorporeal perfusion system to model these effects ex vivo, and combined this with a phospho-CyTOF workflow to comprehensively evaluate single-cell signatures of early activation across all major circulating immune subsets...
July 28, 2017: Journal of Immunological Methods
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