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Traditional measurement of viral titers during vaccine development often involve cytopathic effect (CPE) quantification by plaque assays or tissue culture infectious dose (TCID50) assays, which are based on subjective visual interpretation of cell culture deterioration. These labor-intensive endpoint assays can only provide a mere snapshot of the infection process and the overall kinetics information is inherently missing.
The xCELLigence® Real-Time Cell Analysis technology, a unique live cell assay which allows label-free and continuous tracking of CPE progression in virus-infected cells, provides a dramatically simplified workflow for diverse virology assays, including but not limited to:
- Virus titer determination
- Detection and quantification of neutralizing antibodies
- Antiviral drug discovery & development
- Oncolytic virus characterization
- Quantitative comparison of fitness between different virus strains
Sanofi Pasteur, France
xCELLigence: a new real-time and high-throughput tool for viral infectivity titration
The classical cell-culture methods, such as cell culture infectious dose 50% (CCID50) assays, are time-consuming, end-point assays currently used during the development of a viral vaccine production process to measure viral infectious titers. However, they are not suitable for handling the large number of tests required for high-throughput and large-scale screening analyses. Impedance-based bio-sensing techniques used in real-time cell analysis (RTCA) to assess cell layer biological status in vitro, provide real-time data. In this talk, we will show the assessment of RTCA virus-titration assay as an alternative method to the classical CCID50 assay during the development of viral vaccine production process.
Brandon Lamarche, Ph.D.
Monitoring Virus Cytopathic Effects in Real-Time: Diverse Applications Ranging from Vaccine Development and Drug Discovery to Quantification of Neutralizing Antibodies
When infected by a virus, host cells typically display phenotypic changes (such as swelling, detachment, and lysis) that are collectively referred to as a cytopathic effect (CPE). Although CPEs can be a useful tool for monitoring viral infection, traditional methods like the plaque assay attempt to quantify CPEs manually and are therefore labor intensive, subjective, and suffer from poor reproducibility. Each of these challenges is overcome by the xCELLigence Real-Time Cell Analyzer. Using gold biosensors embedded within the bottom of microtiter plate wells, the xCELLigence instrument continuously monitors virus-induced changes in host cell number, cell size, cell-substrate attachment strength, and cell-cell adhesion (i.e. barrier function). The high sensitivity and reproducibility, simple workflow, and real-time kinetics of this assay make it a powerful tool for diverse virology applications. In this talk I will explain how the xCELLigence biosensors monitor CPEs, discuss the workflow, and will demonstrate the utility of this instrument for virus titer determination, quantification of neutralizing antibodies, antiviral drug discovery/development, evaluating oncolytic virus efficacy and assessing virus quality/fitness.
Date/Time: Wed, Nov 14, 2018 8:00 AM – 9:00 AM PST
Speakers: Brandon Lamarche, Ph.D ACEA Biosciences
Combining the AccuWound 96 with your xCELLigence RTCA instruments gives you better insight into your research by: * Generating 96 identical scratch wounds in seconds * Obtaining true kinetic analysis * Achieving 5x improvement in your coefficient of variation (compared to traditional scratch assays)
Topics covered in this webinar include:
– Scratch assays
– Cell invasion and migration
Date: Thursday, June 28, 2018
Human induced pluripotent stem cell (hiPSC) derived models are heavily explored as alternatives for animal models used in preclinical discovery and safety assessment. Here, Jean-Pierre Valentin from UCB Biopharma will provide a retrospective review on three compounds which showed preclinical and clinical cardiac liabilities. To understand to what extend hiPSC-derived cardiomyocytes could have predicted their side effects, the three compounds were tested using Ncardia’s human iPSC-derived cardiomyocytes in combination with ACEA’s xCELLigence Cardio ECR system. The hiPSC-derived models demonstrated value in refining the integrated cardiac safety assessment.
Using real-time kinetic analysis, this physiologically-relevant human cardiac model allows for the detection of both acute as well as chronic drug-induced toxicities. In the last few years, this predictive validity of hiPSC-derived cardiomyocytes has also been substantiated by various case studies and publications. As a result, pharma are slowly integrating these assays into their early safety assessment to detect potential safety hazards and improve compound selection prior to entering the costly and time-consuming animal studies.
- Uses of iPSC-derived cell types in cardiac screening/safety applications
- Humanizing drug discovery/development, “mimic the clinic”
- ‘Real-world’ examples / case studies
Jean-Pierre Valentin, PhD, HDR, ERT, FSBiol, FRCPath, DSP
Senior Director, Head of Investigative Toxicology
UCB Biopharma SPRL
Jean-Pierre holds a Ph.D. in Physiology & Pharmacology 1990, from the University of Montpellier, France. Following a post-doc at UCSF, Jean-Pierre joined the Pierre Fabre Research Centre (1992-98) where he contributed to the discovery and progression into development of 3 candidate drugs. He joined AstraZeneca to build from inception, develop and lead the Department of Safety Pharmacology where he contributed to the safety evaluation of ~200 candidate drugs across a wide range of therapy areas, leading to the development and successful registration of several marketed products. In February 2014, he joined UCB-Biopharma as Senior Director Head of Investigative Toxicology, based in Belgium. He is a member of several scientific societies; Past President of the Safety Pharmacology Society; co-chair of the HESI subcommittee on QT/Arrhythmia, and the ABPI-NC3Rs-Animal Model Framework. He is involved in training and education programs and is author/co-author of several patents and >180 peer review publications and book chapters.
Xiaoyu Zhang, PhD
Senior Scientist, R & D/Applications
ACEA Biosciences Inc.
Xiaoyu Zhang received her Ph.D. in Cell Molecular and Development Biology from the University of California, Riverside, conducted postdoctoral training and research at Burnham Institute for Medical Research. Dr. Zhang joined ACEA Biosciences in 2011. She has been leading the FDA CiPA studies, cardiac safety screening program and RTCA Cardio/CardioECR applications development.
Greg Luerman, PhD
Technical Director, North America
Dr. Greg Luerman is Technical Director at Ncardia. Following his graduate studies at the University of Louisville School of Medicine, Dr. Luerman earned a Michael J Fox Foundation postdoctoral fellowship within the Pfizer Neuroscience Research Unit where he lead bioassay development on a Parkinson’s disease therapeutic team. He moved on to establish a preclinical cardiac safety/tox and drug discovery assays at ChanTest (now Charles River). Now at Ncardia, Dr. Luerman oversees North American scientific operations.
Dr. Matthew Nystoriak
Assistant Professor of Medicine, Division of Cardiovascular Medicine
University of Louisville
Dr. Matthew Nystoriak is an Assistant Professor of Medicine, Division of Cardiovascular Medicine, and member of the Diabetes and Obesity Center at the University of Louisville. He graduated (PhD) from the Department of Pharmacology at the University of Vermont, College of Medicine, and completed his postdoctoral training in Physiology and Biophysics at the University of Washington in Seattle, and Pharmacology at the University of California, Davis. His current work aims to develop a better understanding of the cross-talk between myocardial energetic demand and coronary microvascular function by elucidating cellular mechanisms coupling intermediary metabolism with vascular ion channel function. In addition, his laboratory is testing how electrical signaling in the cardiovascular system is affected by exposure to chemical constituents of electronic cigarettes.Click here to register
WEBINAR: Measuring the Growth of Microbial Biofilms in Real-Time: Evidence, Insights and Applications
Alex Mira, Ph.D.
Center for Advanced Research in Public Health, FISABIO Foundation, Valencia (Spain)
In this webinar, we will:
- Provide an overview of how impedance-monitoring by xCELLigence instruments can be used to track the dynamics (attachment, growth, dissipation) of bacterial and fungal biofilms
- Validate the utility of the xCELLigence assay by comparing WT vs. mutant strains that are deficient in biofilm production, and by comparing the xCELLigence impedance signal with standard quantification methods such as safranin staining
- Analyze both single- and multi-species biofilms, with the goal of using xCELLigence plates as an in vitro model for complex human-associated biofilms
- Demonstrate the use of xCELLigence for anti-biofilm drug screening and antibiotic susceptibility testing, with comparison to the results obtained by traditional methods such as epsilon-tests
- Look at efforts to employ xCELLigence as a clinical theranostic tool for identifying the optimal treatment regimen for biofilm infections
About the speaker
Alex Mira obtained his Ph.D. in microbiology at Oxford University (UK) in 1999, and carried out post-doctoral research in the USA and Sweden working on microarray technology, microbial genomics, and bioinformatics. In 2003 he started his own research group studying oral microbiota using ‘omics’ techniques. In 2009, he was awarded the “Jaime Ferran” Spanish Award for Research in Microbiology. He is currently the principal investigator of the Oral Microbiome Laboratory at the CSISP, in Spain, where he has applied metagenomics, next-generation sequencing, and impedance technology to study the human microbiome and human-associated biofilms. His team has helped to pioneer use of the xCELLigence instrument for studying microbial biofilms, including applications such as: 1) discovering drugs that inhibit biofilm growth, 2) measuring antibiotic susceptibility in biofilm-mediated infections (i.e. clinical theranostics), and 3) developing an in vitro model of oral diseases.
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are increasingly utilized in basic biology research, drug discovery applications, and for studies of disease mechanisms. The opportunity to measure the contractility, viability, and electrophysiology of hiPSC-CMs in real time over extended periods can provide researchers unique mechanistic insights into the roles cardiomyocytes play in both normal development and cardiac disease.
In this special webinar, scientists using hiPSC-CMs will describe new approaches aimed at enhancing assessment of hiPSC-CM functionality. The speakers will also discuss how these approaches can be used to explore compounds that modulate the force of cardiomyocyte contraction or to assess the safety and efficacy of drugs and drug combinations targeting cardiac tissue.
In this webinar, you will gain insights on:
- New instrumentation to assess cardiomyocyte contractility, viability, and electrophysiology
- Label-free, real-time techniques to assess the “beating” of cardiomyocytes as a biologically-relevant measure of cardiomyocyte function
- Use of electrical pacing for further maturation of cardiomyocytes
- Techniques to detect functional cardiotoxicity
In addition, you will also be able to ask your own specific questions of the speakers during a live question-and-answer session following the presentations.
Armando Lagrutta, PhD
Merck & Co. Inc.
Kristina Green, PhD
Patrick C.H. Lo, PhD
Dan MacLeod Ph.D.
Precision BioSciences, Inc.
Dr. MacLeod received his Ph.D. in Molecular Pathology and Biomedical Sciences from the University of California, San Diego (UCSD). He subsequently conducted postdoctoral research at the International AIDS Vaccine Initiative Neutralizing Antibody Center at the Scripps Research Institute. Dr. MacLeod is currently leading a group in Precision BioScience’s Cell Therapy Research team, focused on expanding the use of Precision’s propriety ARCUS gene-editing technology to engineer T cells for cancer therapy.
Adoptive cellular therapy using chimeric antigen receptor (CAR) T cells has produced significant responses in patients with CD19+ hematological malignancies. Most clinical trials to date have used autologous patient cells, which incur significant manufacturing challenges and costs and are not feasible for many patients with advanced disease state.
At Precision BioSciences, we have developed an allogeneic method using our proprietary ARCUS™ gene-editing technology to generate “universal” CAR T cells from healthy donors. These cells are engineered to eliminate expression of the endogenous T cell receptor making them safe for transfer to unrelated patients. By integrating the CAR transgene at a single genomic site, we can tightly control the level of CAR expression on the cell surface. We further optimize the cells by designing new CAR T constructs that include modifications to the extracellular binding domains as well as the intracellular signaling domains.
Evaluating the impact of these new modifications requires sensitive methods to assay CAR T cell function. Traditional methods for evaluating cytotoxicity cannot effectively predict in vivo activity of a CAR T product; this is because they assess short-term killing potential rather than the ability to expand, persist, and exhibit serial killing. We have used the ACEA xCELLigence instrument for Liquid Tumor Assays to test multiple CAR constructs with modified ectodomains, endodomains, and other modifications, and we have found this platform sensitive enough to detect subtle yet significant differences in performance not observed in traditional killing assays. Furthermore, the sensitivity of the instrument has enabled us to evaluate CAR constructs using cell numbers that are orders of magnitude lower than other assays, greatly enhancing our throughput and capabilities.
For full video, watch above or click here.
Dr. Felix Bohne
Institute of Virology, German Research Center for Environmental Health, Helmholtz Zentrum München
Dr. Sarah K. Lamore
AstraZeneca Pharmaceuticals, Waltham, MA
Dr. Stuart Martin University of Maryland School of Medicine, Baltimore, MDBreast tumor stem cells that are circulating in the bloodstream can invade distant tissues and lie dormant for long periods of time. Reemergence of these disseminated stem cells as metastatic tumors is a primary cause of patient death. In this webinar…
Dr. Stephan Gabos University of Alberta, Edmonton, AB (Canada)Modern public health protection requires ongoing surveillance of environmental media (water, air, food and soil) for potentially harmful chemicals. There is currently an unmet need for in vitro toxicity assays with higher sensitivity, lower interference, and better…
Dr. Sandra N. Garcia Kinetic Concepts Inc., San Antonio, TXNegative Pressure Wound Therapy (NPWT), where a vacuum is applied to an acute or chronic wound, has proven extremely effective for promoting would healing. The success of NPWT lies in its ability to draw the edges of a wound together, to promote granulation,…
Dr. Keith L. Knutson
Mayo Clinic, Jacksonville, FL
The Vaccine & Gene Therapy Institute of Florida, Port Saint Lucie, FL
Dr. Melissa L. Fishel
Indiana University School of Medicine, Indianapolis, IN
Dr. Matthew F. Peters
AstraZeneca Pharmaceuticals, Boston, MA
Powerful Immunotherapy Module at Your Fingertips
RTCA Software Pro is an integrated software package for running and analyzing real time cell analysis data from xCELLigence RTCA DP, SP, and MP instruments. This new version has two modules: RTCA Basic and Immunotherapy. The immunotherapy module is designed to measure cytolysis of target tumor cells by immune effector cells and other molecules. Streamline your experimental design and analysis with this new module.
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For French audio, please click here.