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https://www.readbyqxmd.com/read/29145098/a-novel-simplified-strategy-of-relative-quantification-n-glycan-quantitative-glycomics-using-electrospray-ionization-mass-spectrometry-through-the-stable-isotopic-labeling-by-transglycosylation-reaction-of-mutant-enzyme-endo-m-n175q
#1
Qing Shi, Ryugo Hashimoto, Tadamune Otsubo, Kiyoshi Ikeda, Kenichiro Todoroki, Hajime Mizuno, Dongri Jin, Toshimasa Toyo'oka, Zhe Jiang, Jun Zhe Min
The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. In this study, we describe a novel and simplified quantitative glycomics strategy. Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics...
November 12, 2017: Journal of Pharmaceutical and Biomedical Analysis
https://www.readbyqxmd.com/read/29144195/extraction-purification-of-prostate-specific-antigen-psa-and-establishment-of-radioimmunoassay-system-as-a-diagnostic-tool-for-prostate-disorders
#2
Ahmed Sami Abu-Bakr El-Bayoumy, Akaber Tarek Hessien Keshta, Khaled Mohamed Sallam, Nahed Hassan Ebeid, Hatem Mohamed Elsheikh, Bsheer El-Sayed Bayoumy
This study aimed to provide an easy and effective method for extraction and purification of prostate specific antigen (PSA) from human seminal fluid with high quantity (14 mg) and high purity (98%). The obtained PSA was injected into rabbits for production of anti-PSA polyclonal antibody (titer 1/1000), labeled with radioactive iodine-125 for preparation of radioactive PSA tracer (purity 98 ± 1.8% and specific activity 64 ± 1.9 µCi/µg), and used in preparation of PSA standards. All prepared components can be used in PSA immunoassays specially radioimmunoassay (RIA) kit preparation as a diagnostic tool for prostatic diseases...
November 16, 2017: Journal of Immunoassay & Immunochemistry
https://www.readbyqxmd.com/read/29143881/rmce-based-insect-cell-platform-to-produce-membrane-proteins-captured-on-hiv-1-gag-virus-like-particles
#3
João Vidigal, Bárbara Fernandes, Mafalda M Dias, Marco Patrone, António Roldão, Manuel J T Carrondo, Paula M Alves, Ana P Teixeira
Conformationally complex membrane proteins (MPs) are therapeutic targets in many diseases, but drug discovery has been slowed down by the lack of efficient production tools. Co-expression of MPs with matrix proteins from enveloped viruses is a promising approach to obtain correctly folded proteins at the surface of virus-like particles (VLPs), preserving their native lipidic environment. Here, we implemented a site-specific recombinase-mediated cassette exchange (RMCE) strategy to establish a reusable HIV-1 Gag-expressing insect cell line for fast production of target MPs on the surface of Gag-VLPs...
November 16, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29143683/the-progression-of-heartworm-associated-respiratory-disease-hard-in-spf-cats-18%C3%A2-months-after-dirofilaria-immitis-infection
#4
A Ray Dillon, Bryon L Blagburn, Michael Tillson, William Brawner, Betsy Welles, Calvin Johnson, Russell Cattley, Pat Rynders, Sharron Barney
BACKGROUND: Heartworm-associated respiratory disease (HARD) in cats is induced by the arrival and death of immature adult Dirofilaria immitis in the pulmonary system and is indistinguishable from mature adult heartworm infection. METHODS: A controlled, blind research study investigated the long-term (18 months post infection, PI) consequences of the inflammatory response associated with the death of immature adult heartworms in cats. Three groups of cats, 10 per group, were infected with 100 third-stage (L3) larvae by subcutaneous injection...
November 9, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/29143661/heartworm-associated-respiratory-disease-hard-induced-by-immature-adult-dirofilaria-immitis-in-cats
#5
A Ray Dillon, Byron L Blagburn, Michael Tillson, William Brawner, Betsy Welles, Calvin Johnson, Russell Cattley, Pat Rynders, Sharron Barney
BACKGROUND: A controlled, blind research study was conducted to define the initial inflammatory response and lung damage associated with the death of immature adult Dirofilaria immitis in cats as compared with cats developing adult heartworm infections and cats on preventive medication. METHODS: Three groups of cats were utilized, 10 per group. All cats were infected with 100 third-stage (L3) larvae by subcutaneous injection. Group A cats were treated topically with selamectin (Revolution®; Zoetis) per label directions at 28 days post infection (PI) and once monthly for 8 months...
November 9, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/29142346/conjugation-of-antibodies-with-radiogold-nanoparticles-as-an-effector-targeting-agents-in-radiobioconjugate-cancer-therapy-optimized-labeling-and-biodistribution-results
#6
Pankaj Garg, Daya Kishore Hazra
Purpose of the Study: Drug accessibility to the tumor cells is an important area of concern with an anticipation of increasing the efficacy of the drug to be delivered to a specific site. The biogenesis of gold nanoparticles using plant-mediated phytochemical extracts and their possible linkage to cancer antibodies with an aim at delivering the conjugate specifically to the tumor-associated antigen is the basic objective of the research. Materials and Methodology: Radiolabeling of antibodies with gold nanoparticles was carried out by a protocol, and the labeling extent of antibodies was compared with that of a radiogold solution to ordinary particulate size (AuNO-Ab)...
October 2017: Indian Journal of Nuclear Medicine: IJNM: the Official Journal of the Society of Nuclear Medicine, India
https://www.readbyqxmd.com/read/29141749/visualization-of-west-nile-virus-in-urine-sediment-using-electron-microscopy-and-immunogold-up-to-nine-years-postinfection
#7
Kristy O Murray, Steven Kolodziej, Shannon E Ronca, Rodion Gorchakov, Patricia Navarro, Melissa S Nolan, Amber Podoll, Kevin Finkel, Sreedhar Mandayam
West Nile virus (WNV) is an important emerging flavivirus in North America. Experimental studies in animals infer the development of persistent infection in the kidneys. In humans, recent studies suggest the possibility of persistent renal infection and chronic kidney disease. Considering the discrepancies between published studies on viral RNA detection in urine of convalescing WNV-positive patients, we explored the use of electron microscopy (EM) with anti-WNV E protein antibody immunogold labeling to detect virus in the urine sediment from a subset of study participants in the Houston WNV cohort...
October 23, 2017: American Journal of Tropical Medicine and Hygiene
https://www.readbyqxmd.com/read/29140548/a-phase-ib-open-label-single-arm-study-to-assess-the-safety-pharmacokinetics-and-impact-on-humoral-sensitization-of-sanguinate-infusion-in-patients-with-end-stage-renal-disease
#8
Bassam G Abu Jawdeh, E Steve Woodle, Abbie D Leino, Paul Brailey, Simon Tremblay, Tonya Dorst, Mouhamad H Abdallah, Amit Govil, Daniel Byczkowski, Hemant Misra, Abraham Abuchowski, Rita R Alloway
The endeavor to study desensitization in kidney transplantation hasn't been matched by an effort to investigate strategies to prevent sensitization. In this study (NCT02437422), we investigated the safety, impact on sensitization and pharmacokinetics of SANGUINATE (SG), a hemoglobin-based oxygen carrier, as a potential alternative to packed red blood cells (PRBC) in transplant candidates with end-stage renal disease (ESRD). Ten ESRD subjects meeting inclusion/exclusion (I/E) criteria were planned to receive 3 weekly infusions of SG (320mg/Kg)...
November 15, 2017: Clinical Transplantation
https://www.readbyqxmd.com/read/29140390/a-carboxylated-graphene-nanodisks-glucose-oxidase-nanotags-and-mn-cds-tio2-matrix-based-dual-signal-amplification-strategy-for-ultrasensitive-photoelectrochemical-detection-of-tumor-markers
#9
Jie Song, Shuo Wu, Xinlan Yang, Jingli Yuan
Graphene nanodisks with good conductivity and plenty of edge sites were synthesized to load glucose oxidase (GRD-GOD) and coupled with a Mn(2+) doped CdS quantum dot (QD) modified TiO2 electrode (CdS:Mn/TiO2) for a highly sensitive photoelectrochemical (PEC) immunoassay. The specific immune-recognition behaviour can bring the GRD-GOD labelled antigen into the antibody immobilized CdS:Mn/TiO2 interface and dramatically enhance the photocurrent response via a dual signal amplification strategy. First, graphene nanodisks with a strong electron transfer capacity can improve the conductivity of both the insulating protein layers and the CdS:Mn/TiO2 matrix, thus facilitating the regeneration of trapped carriers and hot electrons in the CdS:Mn QD films and enhancing the PEC performance...
November 15, 2017: Analyst
https://www.readbyqxmd.com/read/29140158/endosomal-trafficking-regulates-receptor-mediated-transcytosis-of-antibodies-across-the-blood-brain-barrier
#10
Arsalan S Haqqani, Christie E Delaney, Eric Brunette, Ewa Baumann, Graham K Farrington, William Sisk, John Eldredge, Wen Ding, Tammy-Lynn Tremblay, Danica B Stanimirovic
Current methods for examining antibody trafficking are either non-quantitative such as immunocytochemistry or require antibody labeling with tracers. We have developed a multiplexed quantitative method for antibody 'tracking' in endosomal compartments of brain endothelial cells. Rat brain endothelial cells were co-incubated with blood-brain barrier (BBB)-crossing FC5, monovalent FC5Fc or bivalent FC5Fc fusion antibodies and control antibodies. Endosomes were separated using sucrose-density gradient ultracentrifugation and analyzed using multiplexed mass spectrometry to simultaneously quantify endosomal markers, receptor-mediated transcytosis (RMT) receptors and the co-incubated antibodies in each fraction...
January 1, 2017: Journal of Cerebral Blood Flow and Metabolism
https://www.readbyqxmd.com/read/29138881/a-harmonized-immunoassay-with-liquid-chromatography-mass-spectrometry-analysis-in-egg-allergen-determination
#11
Masaomi Nimata, Hideki Okada, Kei Kurihara, Tsukasa Sugimoto, Tsutomu Honjoh, Kazuhiko Kuroda, Takeo Yano, Hirofumi Tachibana, Masahiro Shoji
Food allergy is a serious health issue worldwide. Implementing allergen labeling regulations is extremely challenging for regulators, food manufacturers, and analytical kit manufacturers. Here we have developed an "amino acid sequence immunoassay" approach to ELISA. The new ELISA comprises of a monoclonal antibody generated via an analyte specific peptide antigen and sodium lauryl sulfate/sulfite solution. This combination enables the antibody to access the epitope site in unfolded analyte protein. The newly developed ELISA recovered 87...
November 14, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/29137310/ex-vivo-evaluation-of-tumor-cell-specific-drug-responses-in-malignant-pleural-effusions
#12
Carl-Olof Hillerdal, Rita Ötvös, Tünde Szatmári, Sulaf Abd Own, Gunnar Hillerdal, Åsa-Lena Dackland, Katalin Dobra, Anders Hjerpe
The effect of chemotherapy may be improved by combining the most effective drugs based on testing the sensitivity of the individual tumor ex vivo. Such estimations of tumor cells from effusions have so far not been implemented in the clinical routine as a basis for individualized choice of therapy. One obstacle for such analyses is the admixture of benign cells that might obscure the results. In this paper we test and compare two ways of performing the analysis specifically on tumor cells. First we enrich the tumor cells, using antibody labeled magnetic separation, and measure the effects of subsequent drug exposure with the metabolic activity assays WST-1 and alamar blue...
October 10, 2017: Oncotarget
https://www.readbyqxmd.com/read/29137115/cost-effective-and-handmade-paper-based-immunosensing-device-for-electrochemical-detection-of-influenza-virus
#13
Sivaranjani Devarakonda, Renu Singh, Jyoti Bhardwaj, Jaesung Jang
Although many studies concerning the detection of influenza virus have been published, a paper-based, label-free electrochemical immunosensor has never been reported. Here, we present a cost-effective, handmade paper-based immunosensor for label-free electrochemical detection of influenza virus H1N1. This immunosensor was prepared by modifying paper with a spray of hydrophobic silica nanoparticles, and using stencil-printed electrodes. We used a glass vaporizer to spray the hydrophobic silica nanoparticles onto the paper, rendering it super-hydrophobic...
November 11, 2017: Sensors
https://www.readbyqxmd.com/read/29137040/a-multicenter-open-label-phase-iii-study-of-abcertin-in-gaucher-disease
#14
Beom Hee Lee, Ahmed Fathy Abdalla, Jin-Ho Choi, Amal El Beshlawy, Gu-Hwan Kim, Sun Hee Heo, Ahmed Megahed Hassan Megahed, Mona Abdel Latif Elsayed, Tarik El-Sayed Mohammad Barakat, Khaled Mohamed Abd El-Azim Eid, Mona Hassan El-Tagui, Mona Mohamed Hamdy Mahmoud, Ekram Fateen, June-Young Park, Han-Wook Yoo
BACKGROUND: Gaucher disease (GD) is caused by a deficiency in the lysosomal enzyme glucocerebrosidase. Enzyme replacement therapy (ERT) is recommended for clinical improvement. METHODS: The efficacy and safety of a new imiglucerase, Abcertin, were assessed in 7 Egyptian patients with treatment-naïve type 1 GD. Each patient was administered a biweekly 60 U/kg dose of Abcertin for 6 months. The primary endpoint was the change in hemoglobin concentration. The secondary endpoints were changes from baseline in platelet counts, spleen and liver volumes, biomarker levels, skeletal parameters, and bone mineral density...
November 2017: Medicine (Baltimore)
https://www.readbyqxmd.com/read/29136915/a-quantum-dot-based-lateral-flow-immunoassay-for-the-sensitive-detection-of-human-heart-fatty-acid-binding-protein-hfabp-in-human-serum
#15
Mihaela Savin, Carmen-Marinela Mihailescu, Iulia Matei, Dana Stan, Carmen Aura Moldovan, Marian Ion, Ion Baciu
We describe the preparation and validation of a novel lateral flow immunoassay test for the detection of human heart fatty acid binding protein (hFABP). Water-soluble CdTe quantum dots (QDs) were selected as the fluorescent label and were linked covalently to anti-hFABP antibodies. Upon conjugation, the secondary structure of the anti-hFABP was preserved and the fluorescence quantum yield of the CdTe QDs increased. The labelled antibodies were transferred to the immunoassay test strip and the antigen-antibody reaction was successfully performed...
February 1, 2018: Talanta
https://www.readbyqxmd.com/read/29136858/monoclonal-antibody-n-glycosylation-profiling-using-capillary-electrophoresis-mass-spectrometry-assessment-and-method-validation
#16
Jérémie Giorgetti, Valentina D'Atri, Julie Canonge, Antony Lechner, Davy Guillarme, Olivier Colas, Elsa Wagner-Rousset, Alain Beck, Emmanuelle Leize-Wagner, Yannis-Nicolas François
Characterization of therapeutic proteins represents a major challenge for analytical sciences due to their heterogeneity caused by post-translational modifications (PTM). Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). As a consequence, glycosylation profiling must be deeply characterized. For this application, several analytical methods such as separation-based or MS-based methods, were evaluated...
February 1, 2018: Talanta
https://www.readbyqxmd.com/read/29136590/ultrasensitive-electrochemical-immunosensor-for-quantitative-detection-of-hbeag-using-au-pd-mos2-mwcnts-nanocomposite-as-enzyme-mimetic-labels
#17
Zengqiang Gao, Yueyun Li, Xiaobo Zhang, Jinhui Feng, Ling Kong, Ping Wang, Zhiwei Chen, Yunhui Dong, Qin Wei
A sensitive sandwich-type electrochemical immunosensor for the detection of hepatitis B e antigen (HBeAg) was successfully developed based on the gold@palladium nanoparticles (Au@Pd NPs) loaded by molybdenum disulfide functionalized multiwalled carbon nanotubes (Au@Pd/MoS2@MWCNTs). The resultant nanocomposites not only possessed high specific surface area and good biocompatibility, but also exhibited excellent electro-catalytical property. Au NPs functionalized porous graphene oxide (p-GO@Au) were used as sensing platforms and primary antibodies carriers, which can accelerate the electron transfer and improve the load capacity of primary antibodies (Ab1), improving the sensitivity of the immunosensor...
November 9, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/29134440/ki67-labeling-index-assessment-and-prognostic-role-in-gastroenteropancreatic-neuroendocrine-neoplasms
#18
REVIEW
Günter Klöppel, Stefano La Rosa
In 1983, a monoclonal antibody, Ki67, was generated, that labeled the nuclei of proliferating non-neoplastic and neoplastic cells. The name Ki67 derived from the city of Kiel (Ki) where the antibody was produced in the university department of pathology and refers to the number of the original clone (67). Systematic assessment of the proliferative activity of tumors using Ki67 started in the 1990s, when Ki67, which only worked on frozen tissue, was complemented by the antibody MIB-1 that also worked in formalin-fixed tissues...
November 13, 2017: Virchows Archiv: An International Journal of Pathology
https://www.readbyqxmd.com/read/29134300/freeze-fracture-new-avenues-for-the-ultrastructural-analysis-of-cells-in-vitro
#19
REVIEW
Carola Meier, Anja Beckmann
The ultrastructural analysis of biological membranes by freeze fracture has a 60-year tradition. In this review, we summarize the benefits of the freeze-fracture technique and review special structures analyzed by freeze fracture and by combined freeze-fracture replica immunogold labeling (FRIL) of cell cultures. In principle, every cellular membrane whether of cell suspensions, mono- or bilayers of cell cultures can be analyzed in freeze fracture. The combination of freeze fracture and immunogold labeling of the replica allows the ultrastructural identification of protein assemblies in combination with the molecular identification of their constituent proteins using specific antibodies...
November 13, 2017: Histochemistry and Cell Biology
https://www.readbyqxmd.com/read/29133712/meningiomas-objective-assessment-of-proliferative-indices-by-immunohistochemistry-and-automated-counting-method
#20
Pooja Chavali, Megha S Uppin, Shantveer G Uppin, Sundaram Challa
BACKGROUND: The most reliable histological correlate of recurrence risk in meningiomas is increased mitotic activity. Proliferative index with Ki-67 immunostaining is a helpful adjunct to manual counting. However, both show considerable inter-observer variability. A new immunohistochemical method for counting mitotic figures, using antibody against the phosphohistone H3 (PHH3) protein was introduced. Similarly, a computer based automated counting for Ki-67 labelling index (LI) is available...
November 2017: Neurology India
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