J Control Release. . Sunju Choi, Hittu Matta, Ramakrishnan Gopalakrishnan, Venkatesh Natarajan, Songjie Gong, Alberto Jeronimo, Wei-Ying Kuo, Bryant Bravo, Preet M. Chaudhary> ;Scientific Reports. Photochemical & Photobiological Sciences 2019, 18 (8) , 2061-2070. https://doi.org/10.1039/C9PP00174C, Felix Weihs, Helen Dacres. The sensitivity and signal-to-background ratio were improved considerably by developing a carboxy-terminal fragment engineered from a click beetle luciferase. FIELD. In vivo use of monoclonal antibodies has become routine clinical practice in the treatment of human cancer. A novel thermostable beetle luciferase based cytotoxicity assay, Scientific Reports April 26, 2021, See publication, Development of theranostic active-targeting boron-containing gold nanoparticles. . November 29, 2017. Introduction. 1991, Schafer, Schafer et al. reported a novel BRET-based biosensor for . Using luciferase and fluorescent proteins as reporters, various BL imaging systems have been developed innovatively for real-time monitoring of diverse molecules in vivo based on the reaction between luciferin and the substrate. [ MEDLINE Abstract ] A retrospective matched cohort study evaluating the effects of percutaneous endoscopic gastrostomy feeding tubes on nutritional status and survival in . CD38 is an attractive target, because it has double roles, as a receptor and an ectoenzyme. Growth . Biophotonic cytotoxicity assay for high-throughput screening of cytolytic killing. Sunju Choi, Hittu Matta, Ramakrishnan Gopalakrishnan, Venkatesh Natarajan, Songjie Gong, Alberto Jeronimo, Wei-Ying Kuo, Bryant. A novel thermostable beetle luciferase based cytotoxicity assay. The basic mechanism of luciferases involves the oxidation of the substrate luciferin with the formation of energy-rich peroxy intermediates whose spontaneous decomposition yields singlet excited oxyluciferin which decays emitting light with high efficiency [ 24, 25 ]. Choi S, Matta H, Gopalakrishnan R, Natarajan V, Gong S, Jeronimo A, Kuo WY, Bravo B, Chaudhary PM. Full Text More Novel Immunotherapies sentence examples. MetRS inhibitors, as validated by Compound-1717, have promising anti-giardiasis properties that merit further study as alternative therapeutics. A real time Metridia luciferase based non-invasive reporter assay of mammalian cell viability and cytotoxicity via the -actin promoter and enhancer, PLoS ONE, (2012) P. Wu et al. 1997). Bright luminescence with a high signal-to-background ratio, ideally red or near-infrared light as the emission maximum, is necessary for in vivo animal experiments. We recently described a. A novel thermostable beetle luciferase based cytotoxicity assay. The Caspase-Glo 3/7 assay provides a proluminescent caspase-3/7 DEVD-aminoluciferin substrate and a proprietary thermostable luciferase in a reagent optimized for caspase-3/7 activity, luciferase activity and cell lysis. Development and characterization of a novel luciferase based cytotoxicity assay. Each issue is devoted to a single major therapeutic area guest edited by an acknowledged authority in the field. By coupling isotopic labeling and a novel pairing algorithm, our approach performs an in silico enrichment of acetyl peptides, circumventing the need for immunoenrichment. Based on the the results, a mixture of DMSO and ethanol was superior to the 66.7 % ethanol in the Tricine solution for producing both signal and noise. Although the library of the compounds showed a considerable acute cytotoxicity, a genotoxicity could not be observed. We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives. Go to: Introduction, Here, we report a more than doubling of the maximum bioluminescence wavelength range using a single synthetic . Development of a simple new flow cytometric antibody-dependent cellular cytotoxicity (ADCC) assay with excellent sensitivity . red-shifted FPs as BRET acceptors for BRET-based imaging in vitro and in vivo. our models are very simple and include novel physically-based descriptors, better allowing for large-scale searches of . In this study, we describe the development of a new cytotoxicity assay termed 'Matador-Glo assay' which takes advantage of a thermostable variant of Click Beetle Luciferase (Luc146-1H2). 10 AkaLumine (a d -luciferin analogue that is not recognized by the CBG99 enzyme) was used as a control. beetle red (CBRed) and click beetle green (CBGr68 and CBGr99) [56-59]. In this study, we describe the development of a new cytotoxicity assay termed `Matador-Glo assay' which takes advantage of a thermostable variant of Click Beetle Luciferase (Luc146-1H2). Choi S, Matta H, Gopalakrishnan R, Natarajan V, Gong S, Jeronimo A, Kuo WY, Bravo B, Chaudhary PM. This screening strategy is the simplest among reported in the literature and can potentially be used to efficiently increase thermostability of other beetle or non-beetle luciferases. There is also an active search to produce stable, red-shifted mutants of Renilla [55]. A luciferase release-based cytotoxicity assay was first described by Schafer et al. In this study, we systematically investigated the toxicity of AgNPs exposure using Drosophila melanogaster. A luciferase release-based cytotoxicity assay was first described by Schafer et al in 1997 using Fluc (Schafer, Schafer et al. . . Various attempts have . reported a click beetle luciferase mutant . We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives. 1997). 102:169-77. Measurement of Cytotoxicity by ATP-based Luminescence Assay in Primary Cell Cultures and; Cell Lines. The recoveries of CAs added to rat brain sample were more than 94.6%, with the precisions of less than 5.5%. Each thematic issue of Current Pharmaceutical . Two main approaches for T-cell redirection involve their genetic modification with chimeric antigen receptors (CAR), or the use of recombinant proteins designated bispecific T-cell engagers (BiTE). Adding the single Caspase-Glo 3/7 Reagent in an "add-mix-measure" format results in cell lysis, followed by caspase cleavage of the substrate. In the past decades, the firefly luciferin-luciferase system has been used in an enormous variety of bioanalytical applications. Arne Lundin studies Cognitive Ageing, Categorization, and Research Journal of Medical Sciences Human Physiology Rural Health Science and Mathematics Education. Biochem J. A functional analysis of the CREB signaling pathway using HaloCHIP-chip and high throughput reporter assays. Read the latest Research articles in Health sciences from Scientific Reports We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives. 17,18 Plates were incubated for time increments between 5 and 180 min at room temperature. by Christine Brown. The Promega mission statement is: To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide. Jun 2019 - Jun 20201 year 1 month. Single-cell transfection technologies for cell therapies and gene editing. Since the bioluminescence is detected before and after the heating step it makes a decrease in activity unlikely for the selected mutants. In the last decades bioluminescent reporters have been employed extensively for the development of cell-based assays and in vivo imaging -.Molecular aspects of color variation in luciferase emission have been the subject of numerous studies: various mechanisms have been proposed and multicolor reporter proteins have been created and applied in cell-based assays -. The activities of the luciferases can be measured simultaneously and quantitatively with optical filters. A. Firefly luciferase genes have been widely used as reporters in gene expression studies and for cell tracking in normal biological and pathological processes, including cell proliferation studies, cytotoxicity assays, and metastasis in model animals, emerging as a . Under the optimized conditions, the detection limits defined as a signal-to-noise ratio of 3, were in the range of 1.32-1.90 ng/mL, corresponding to 26.4 - 38.0 pg for 20 L sample injection. Creation of a thermostable firefly luciferase with pH . A novel thermostable beetle luciferase based cytotoxicity assay. A thermostable beetle luciferase that retains at least 50% activity in aqueous solution for at least 2 hours at a temperature of about 50 C., which thermostable beetle luciferase comprises at least one amino acid substitution relative to a reference beetle luciferase, and which comprises SEQ ID NO:14, SEQ ID NO:19, SEQ ID NO:24, SEQ ID NO:26 . DOI: 10.1016/j.jes.2021.12.027, 6. ApoTox-Glo Triplex Assay is a novel multiplex analysis system. Research Interests: Functional Analysis, Genomics, Signal Transduction, Biological Sciences, DNA, and 9 more. . In particular, the . [ { "pmid": "28236482", "articlepubdate": "20170515", "articletitle": "Cytotoxicity and molecular effects of biocidal disinfectants (quaternary ammonia . A Simple and Sensitive Method for Measuring Tumor-Specific T Cell Cytotoxicity. Description. In this study, we describe the development of a new cytotoxicity assay termed 'Matador-Glo assay' which takes advantage of a thermostable variant of Click Beetle Luciferase (Luc146-1H2). Chief, Nohl Division of Hematology and Center for Blood Diseases in the Department of Medicine, Director for Bone Marrow Transplant, Medicine, 1441 Eastlake Ave. Health Sciences Campus Los Angeles, Contact, +1 323 865 3950, pchaudha@usc.edu, Overview, Awards, Publications, Development and characterization of a novel luciferase based cytotoxicity assay. Cree, I. Dr. Matta's research interests include KSHV-associated malignancies and drug development. Meanwhile, the kinetics of protein interactions even in deep tissues has been studied by BL imaging. A., and P. E. Andreotti. J-GLOBAL is a service based on the concept of Linking, Expanding, and Sparking, linking science and technology information which hitherto stood alone to support the generation of ideas.By linking the information entered, we provide opportunities to make unexpected discoveries and obtain knowledge from dissimilar fields from high-quality science and technology information within and outside JST. Madison, Wisconsin Area. The nematodes' preference of the beetle hosts may be explained by the differences in beetle chemical profiles and species-specific microbiome (the aggregate of microorganisms that exist on the surface and inside a host). Finally, the triazene 14 was selected with the best biological properties (IC50 = 3.26 M, NI50 = 24.22 M, SI = 7.44). Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.) A novel thermostable beetle luciferase based cytotoxicity assay. 1998. Sci Rep. 2021 05 11; 11(1):10002. using Fluc 8. 2021;330: 963 - 975. These results were produced from a run with 3000 clusters. Membrane-permeable luciferin esters for assay of firefly luciferase in live intact cells. A new luciferase isolated from Luciola italica has been recently cloned, and thermostable red- and green-emitting mutants were obtained by random and site-directed mutagenesis. November 21, 2014 [ MEDLINE Abstract ] Publication Date: 2009. A novel thermostable beetle luciferase based cytotoxicity assay Here, we outline an open-source semi-automated pipeline to The new in vitro and in vivo assays based on luciferase expression by engineered G. lamblia strains are useful for the discovery and development of new therapeutics for giardiasis. Methods Mol Biol. . Sci Rep. 2021 05 11; 11(1):10002. Provided herein are methods for the real-time monitoring of an event or response. Yoga instructor: I coach my students to develop their mind-body connection using various aspects of yoga including Vinyasa flow . We demonstrate its utility in spatiotemporal characterization of Smad1-Smad4 and Smad2-Smad4 interactions in early developing stages of a single living Xenopus laevis embryo. | Find, read and cite all the research you . We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives. A highly efficient, thermostable and cadmium selective firefly luciferase suitable for ratiometric metal and pH biosensing and for sensitive ATP assays. These novel observations are described in detail in this paper. +1 323 865 3950, hmatta@usc.edu, Overview, Publications, Dr. Matta earned her Master's and PhD in Microbiology followed by a fellowship at The Hamon Center for Therapeutic and Oncology Research, University of Texas Southwestern Medical Center. We show here that AgNPs significantly de . Luminescence-based cell viability testing. part of the assay, cell . We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives. Expired - Fee Related, expires 2025-05-03 Application number US10/954,840 Other versions Sci Rep. 2018 01 09; 8(1):199. 2021 May 11, 8 citations, The time to offer treatments for COVID-19. In this study, we describe the development of a. In vivo bioluminescence imaging (BLI), which is based on luminescence emitted by the luciferase-luciferin reaction, has enabled continuous monitoring of various biochemical processes in living animals. MEY-1 with stability over a broad pH range and significant starch hydrolysis capacity. The CellTiter-Glo Assay relies on the properties of a proprietary thermostable luciferase (Ultra-Glo Recombinant Luciferase), which generates a stable glow-type luminescent signal and improves performance across a wide range of assay conditions. A novel thermostable beetle luciferase based cytotoxicity assay. and particularly robust Ultra-Glo Luciferase, . Cytotoxicity assays are essential for the testing and development of novel immunotherapies for the treatment of cancer. To stimulate natural product-based drug discovery in both the intramural and extramural research community, the NCI Program for Natural Product Discovery has developed automated, high-throughput methods for the generation of an HTS-amenable library of 1,000,000 natural product fractions, which will be made available, free of charge, to researchers in 384-well plates for screening against any . The aminoluciferin serves as a substrate for the thermostable. The microtiter well-based binding assay showed to be over 16-fold more sensitive in detecting Gluc in the blood as compared to the direct standard approach showing that our optimized protocol . 1997. Download Free PDF Download PDF Download Free PDF View PDF. . The optimum concentration of luminescent substrate, d -luciferin (GoldBio, USA), required for cell identification was determined as described earlier. Coleopteran bioluminescence is unique in that beetle luciferases emit colors ranging between green (ca.550 nm) and red (ca.600 nm), including intermediate colors such as yellow and orange, allowing up to 3 simultaneous parameters to be resolved in vitro with natural luciferin (D-LH2). In the first. Article Title: A novel thermostable beetle luciferase based cytotoxicity assay Journal: Scientific Reports doi: 10.1038/s41598-021-89404-z However, the shorter half-life of Fluc (<30 minutes) in tissue culture medium hindered its wider use (Thompson, Hayes et al. Different combinations of luciferases were used in vitro as purified proteins and expressed in bacterial and mammalian cells to test their suitability for multicolor assays. Sci Rep. 2021;11: 10002. , , [Web of Science ], [Google Scholar] Duckert B, Vinkx S, Braeken D, et al. [16] Cluster path (and word type count) Words (most frequent) ^00000000 (53) changes alterations ^0000000100 (41) variation variations fluctuations imbalances divergence dependencies ^0000000101 (79) variability fluctuation variances turbulence ^0000000110 (62) trends savings gaps disparities saving inequalities inconsistencies tendencies . In this study, we describe the development of a new cytotoxicity assay termed 'MatadorGlo assay' which takes advantage of a thermostable variant of Click Beetle Luciferase (Luc1461H2). 10.3390/cancers13061363. for determining viability, cytotoxicity and apoptosis of cell lines . Promega Corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, high-value products for the life sciences. In this study, we describe the development of a new cytotoxicity assay termed 'Matador-Glo assay' which takes advantage of a thermostable variant of Click Beetle Luciferase (Luc146-1H2). Cree, I. The luciferase reaction for this assay is shown in Figure 3. We have developed a novel reporter assay system in which three luciferases that emit green, orange, and red light with a single substrate are used as reporter genes. A novel thermostable beetle luciferase based cytotoxicity assay. -gp130 and V(L)-EpoR) were expressed in an IL-3-dependent myeloid cell line, 32D. PDF | Cytotoxicity assays are essential for the testing and development of novel immunotherapies for the treatment of cancer. Comparison of red-shifted firefly luciferase Ppy RE9 and conventional Luc2 as bioluminescence imaging reporter genes for <italic>in vivo</italic> imaging of stem cells. A novel approach for detecting viable and tissue-specific circulating tumor cells through an adenovirus-based reporter vector, Prostate, (2014) Here, we developed and employed a novel method for directly quantifying stoichiometry of site-specific acetylation in the entire proteome of Escherichia coli. The presence of a continuous output signal that can represent realtime metabolic activity dynamics offers enhanced temporal resolution for assaying the cytotoxicity of therapeutic compounds in high throughput screening formats and provides for standardization without the need to tailor each assay to the unknown kinetics of novel compounds [ 19 ]. AgNPs exposure have been reported to cause toxicity in animals. A novel triple-modality reporter gene for whole-body fluorescent, bioluminescent, and . However, the shorter half-life of Fluc (<30 minutes) in tissue culture medium hindered its wider. by Jacqui Mndez and Keith V Wood. Aims & Scope Current Pharmaceutical Design publishes timely in-depth reviews/ mini reviews and research articles from leading pharmaceutical researchers in the field, covering all aspects of current research in rational drug design. Development of luciferase-linked antibody capture assay based on luciferase . A novel thermostable beetle luciferase based cytotoxicity assay. We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives. Development of a Novel Nonradioisotopic Assay and Cdc25B Overexpression Cell Lines for Use in Screening for Cdc25B Inhibitors . luciferase seq wild type amino acid cnbluc03 Prior art date 2003-10-03 Legal status (The legal status is an assumption and is not a legal conclusion. firefly luciferase, luciferin, photinus pyralis luciferase, beetle luciferase, pc3-luc, pplase, cbrluc, luciola italica luciferase, lucppy, ppy re-ts, more top print hide show all columns Go to Synonym Search In this study, we describe the development of a new cytotoxicity assay termed 'Matador-Glo assay' which takes advantage of a thermostable variant of Click Beetle Luciferase (Luc146-1H2). The mixture of dimethyl sulfoxide (DMSO) and ethyl alcohol (9;1, v/v) and 66.7 % ethanol in the Tricine solution (pH 12.5) were examined for producing peroxy BSA. Sci Rep. 2018 01 09; 8(1):199. Improved luciferase-based assays WO2004081189A2 (en) 2003-03-07: 2004-09-23: . Publication Name: BMC Genomics. Novel anti-CD38 humanized mAb SG003 possessed enhanced cytotoxicity in lymphoma than Daratumumab via antibody-dependent cell-mediated cytotoxicity. 276:637-41. A novel thermostable beetle luciferase based cytotoxicity assay. Visualization and quantitation of cellular cytotoxicity using cell-permeable fluorogenic protease substrates and caspase activity indicator markers . In this study, we describe the development of a. Cytotoxicity Assay and the Caspase-Glo 3/7 Assay. Hall et al. A modified beetle luciferase protein which is an environmentally sensitive reporter protein is provided. We recent-ly described a novel cytotoxicity assay, termed the Matador as-say, . . by Bryant Bravo. Thermostable red and green firefly luciferase mutants are also being optimized for dual-color bioluminescent reporter assays potentially A thermostable glucoamylase from Bispora sp. renaturation of the immobilized luciferase since the renaturation yield decreased at acidic pH only for the immobilized luciferase. Cytotoxicity assays are essential for the testing and development of novel immunotherapies for the treatment of cancer. We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives. These approaches have demonstrated dramatic effects in patients with hematologic cancers, although limited effect against solid cancers. Nevertheless, the known mechanisms of AgNPs toxicity are still limited. Hittu Matta's 72 research works with 2,059 citations and 4,476 reads, including: A novel thermostable beetle luciferase based cytotoxicity assay E. Kamrani, R. Stevanato, S. M. Etezad and K. Khajeh, Purification and characterization of a novel thermostable luciferase from . Patent History. A novel thermostable beetle luciferase based cytotoxicity assay, Authors, Sunju Choi # 1 , Hittu Matta # 1 , Ramakrishnan Gopalakrishnan # 1 , Venkatesh Natarajan 1 , Songjie Gong 1 , Alberto Jeronimo 1 , Wei-Ying Kuo 1 , Bryant Bravo 1 , Preet M Chaudhary 2, Affiliations, by Bryant Bravo. 14,84,85,117 Costoya et al. We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives.