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Merck KGaA, Darmstadt, Germany Webinars

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  • Recent developments in RAFT agents
    Recent developments in RAFT agents Dr. James Gardiner-Senior Research Scientist, Dr. Melissa Skidmore-Senior Research Scientist, Dr. Graeme Moad (CSIRO) Recorded: Oct 11 2017 41 mins
    This webinar will provide an overview of recent advances in RAFT agents (900150, 900157 and 900158). New dithiocarbamate RAFT agents are extremely versatile, RAFT agents with wide-spread monomer applicability. The RAFT agents have the distinct advantage of low odour levels and in addition to this and the derived polymers do not develop odour on storage as no low molar mass thiols are generated. In most cases they are an appropriate replacement for trithiocarbonate RAFT agents. The new RAFT agents have the ability to control polymerization of both MAMs (more activated monomers) and LAMs (less activated monomers) and have been shown to be suitable for the synthesis of poly(MAM)-block-poly(LAM), specifically poly(DMA)-blockpoly(VAc).
  • Stable Isotope-Labeled Protein Internal Standards
    Stable Isotope-Labeled Protein Internal Standards James Walters, Ph.D. Recorded: Sep 21 2017 62 mins
    Mass spectrometry-based protein assays impart increased specificity and more rapid development times versus traditional methods, such as ELISA. Coupled with immunoaffinity enrichment, LC-MS/MS is becoming a powerful tool for the quantitation of proteins in plasma. Such methods typically rely on synthetic stable isotope labeled (SIL) peptide internal standards to correct for instrumental variability. For more accurate protein quantitation by LC-MS/MS, experimental variations throughout the entire sample preparation workflow, including protein fractionation, immunoaffinity enrichment, and enzymatic digestion, must be accounted for. An ideal way of improving assay reproducibility is to add a full-length stable isotope labeled recombinant protein, that is equivalent to the native target protein, to the sample at the initial stage of the assay workflow. We have developed a set of stable-isotope-labeled monoclonal antibodies expressed in CHO cells as well as SIL versions of several clinically-relevant human proteins expressed in E. coli, such as IGF1, and in mammalian HEK293 cells, such as Thyroglobulin (manufactured as a Certified Reference Material). We will present data to demonstrate that the use of full-length SIL proteins and antibodies as internal standards allows for more accurate and rapid quantitation of biotherapeutic antibodies and clinically-relevant human protein biomarkers in plasma by LC-MS/MS.
  • Stable Isotope-Labeled Protein Internal Standards
    Stable Isotope-Labeled Protein Internal Standards James Walters, Ph.D. Recorded: Sep 20 2017 56 mins
    Mass spectrometry-based protein assays impart increased specificity and more rapid development times versus traditional methods, such as ELISA. Coupled with immunoaffinity enrichment, LC-MS/MS is becoming a powerful tool for the quantitation of proteins in plasma. Such methods typically rely on synthetic stable isotope labeled (SIL) peptide internal standards to correct for instrumental variability. For more accurate protein quantitation by LC-MS/MS, experimental variations throughout the entire sample preparation workflow, including protein fractionation, immunoaffinity enrichment, and enzymatic digestion, must be accounted for. An ideal way of improving assay reproducibility is to add a full-length stable isotope labeled recombinant protein, that is equivalent to the native target protein, to the sample at the initial stage of the assay workflow. We have developed a set of stable-isotope-labeled monoclonal antibodies expressed in CHO cells as well as SIL versions of several clinically-relevant human proteins expressed in E. coli, such as IGF1, and in mammalian HEK293 cells, such as Thyroglobulin (manufactured as a Certified Reference Material). We will present data to demonstrate that the use of full-length SIL proteins and antibodies as internal standards allows for more accurate and rapid quantitation of biotherapeutic antibodies and clinically-relevant human protein biomarkers in plasma by LC-MS/MS.
  • Guava Flow Cytometry and Duolink
    Guava Flow Cytometry and Duolink Katherine Gillis and Tracy Adair-Kirk Recorded: Aug 10 2017 42 mins
    In this webinar you will learn about DuoLink PLA and the Guava easyCyte flow cytometry systems
  • 3D Nephrotoxicity applications & Nortis’ microfluidic organ-on-chip technology
    3D Nephrotoxicity applications & Nortis’ microfluidic organ-on-chip technology Dr. Ed Kelly, Dr. Henning Mann Recorded: Jul 25 2017 97 mins
    1. The kidney proximal tubule is the primary site of drug-induced nephrotoxicity. I will describe the development of a 3-dimensional flow-directed proximal tubule microphysiological system (MPS). The kidney MPS recapitulates the synthetic, metabolic and transport activities of kidney proximal tubule cells. This MPS is as an ideal platform for ex vivo modeling of nephrotoxicity. Towards this goal, we have evaluated nephrotoxicity in response to challenge with multiple toxicants, including the heavy metal pollutant cadmium, antisense oligonucleotides, the antibiotic polymyxin B and the Chinese herbal product aristolochic acid. We believe that MPS technologies will have major impacts on predictive toxicity testing and human risk assessment. Animal and in vitro systems do not always faithfully recapitulate drug and xenobiotic responses in the clinic or occupational/environmental exposures, respectively. MPS technologies will refine safety assessment and reduce our need for surrogate animal testing. An ultimate goal is to create integrated human MPS organ systems that could replace animal models.

    2. Nortis has developed a technology that is used to recapitulate functional units of human organs in microfluidic devices (chips). Such organ models include vasculature, kidney, and liver models for toxicology studies, blood-brain barrier models for drug transport studies, and vascularized tumor microenvironment models for drug efficacy studies.
  • Developing Simplified Methods for Quantifying Analytes using  SPME
    Developing Simplified Methods for Quantifying Analytes using SPME Robert E. Shirey, M.S.; Principal R&D Scientist Recorded: Jul 13 2017 75 mins
    Solid phase microexatraction or SPME is a green method for extraction of analytes out of a sample. Since SPME is a non-exhaustive extraction technique, some analysts believe that SPME is not quantifiable. This presentation will provide basic information for developing a method to extract and quantify analytes using SPME. Examples will be given on the extraction and quantification of analytes out of various matrices, and SPME will be compared to other extraction techniques such as QuEChERS and SPE. In this webinar, we will discuss some new SPME technologies such as SPME-OC (over-coated) fibers and BioSPME that help to isolate and quantify analytes from interfering compounds in the matrix. Guidelines will be provided for enhancement of precision using SPME.
  • Advanced Cell Culture Technology for Generation of In Vivo-like Tissue Models
    Advanced Cell Culture Technology for Generation of In Vivo-like Tissue Models Stefan Przyborski, PhD - Professor of Cell Technology, Durham University UK Recorded: Jun 30 2017 64 mins
    The benefits of three dimensional (3D) cell culture are widely appreciated. More cell-based technologies are now becoming available that enable researchers to preserve the native 3D structure of cells in vitro. These can be broadly divided into three areas: aggregate-based methods; hydrogels and extra-cellular matrices; and inert scaffold-based technologies. Each has strengths and weaknesses and there is no one technology that satisfies all applications. Tissues in the body are mostly composed of different cell types that are often highly organized in relation to each other. Often cells are arranged in distinct layers that enable signalling and cell-to-cell interactions. Alternatively in tumours, cancer cells form aggregates and tissue masses composed of different cell types. Recreation of these types of architecture will significantly evolve 3D cell culture to a new level where real tissue-like structures can be generated in vitro.

    This webinar will review the alternative approaches available to researchers and provide an overview of their capabilities and example applications. More sophisticated models are developing as 3D cell culture technology becomes established and accepted as a means of creating more physiologically relevant cell-based assays. Methods that are relatively straightforward to use and that recreate the organized structure of real tissues will become valuable research tools for use in discovery, validation studies, and modelling disease.

    Key areas covered:
    • 2D vs 3D cell culture debate
    • Review of alternative approaches and the development of new technologies
    • Challenges facing 3D culture methods, in terms of technologies available and methods used
    • Showcase applications where 3D technology makes a difference
    • Future perspective for 3D cell culture technology and further development
  • Winning Westerns: Proven Strategies to Optimize Your Western Blots
    Winning Westerns: Proven Strategies to Optimize Your Western Blots Natasha L. Pirman, Ph.D. Recorded: Jun 21 2017 53 mins
    Does western blotting give you more trouble than expected? Do you feel like your precious samples are being wasted on bad westerns? Join us and find out how you can improve your western blots! In this seminar, you will learn general guidelines for performing and troubleshooting your westerns, such as:

    • Choice of different blotting membranes
    • Parameters affecting blotting efficiency
    • Conditions for optimizing your immunodetection
    • Information on SNAP i.d.® 2.0 system: A faster way to perform immunodetection
  • Amplified Detection of Proteins and their Interactions using Duolink PLA
    Amplified Detection of Proteins and their Interactions using Duolink PLA Manpreet Mutneja, Ph.D, MBA Sr. Product Manager, Molecular Platforms Life Science Research, MilliporeSigma Recorded: Jun 15 2017 44 mins
    Understanding the movements, modifications and interactions of proteins within a cell is key to unraveling the fundamental tenets of biology. However, the low-level expression of many proteins, combined with the transient nature of their interactions and movements, makes analyzing and understanding these processes quite difficult. Duolink® PLA, which is based on the principles of the proximity ligation assay (PLA), offers a solution to overcome these hurdles and to study the actions of endogenous proteins within cells and tissues. Combining the specificity of antibodies with the sensitivity afforded by rolling circle amplification, Duolink® PLA allows you to detect, visualize, and quantitate proteins and their interactions (even single events) where they happen within cells or tissue, all without overexpression or genetic manipulation. This seminar will cover the basic assay principle and advantages of the Duolink® PLA technology, and discuss recent applications and developments of the technology that make it an excellent tool to understand the fundamental mechanisms of biology, as well as disease states. Applications of Duolink® PLA include the investigation of cellular responses to varying stimuli, receptor dimerization and signalling cascades, post-translational modifications, and regulation of protein expression. New developments include use in flow cytometry and multiplexed detection.
  • Basic Principles of Solid Phase Microextraction (SPME) Method Development
    Basic Principles of Solid Phase Microextraction (SPME) Method Development Emanuela Gionfriddo, Ph.D.; Research Associate under Prof. Janusz Pawliszyn, University of Waterloo (ON, Canada) Recorded: May 31 2017 74 mins
    For the past two decades, Solid Phase Microextraction (SPME) has represented a convenient alternative to conventional sample prep procedures. SPME allows the simultaneous extraction and enrichment of analytes of interest from a given matrix in a single step while avoiding, or drastically minimizing, the use of organic solvents and time-consuming cleanup procedures.
    Like any other analytical method, the various parameters governing the SPME process need to be carefully optimized in order to achieve robustness and sensitivity. However, certain aspects of SPME method development are often overlooked by many users, leading to unsatisfactory performance of the technique.
    This webinar will shed light into several aspects of SPME method development. The presentation will include a theoretical explanation of SPME fundamentals and practical suggestions to overcome common errors and bias encountered when using SPME.
    The webinar is divided in three main sections: 1) optimization of extraction conditions 2) matrix modifications 3) optimization of desorption conditions for gas and liquid chromatography. Each section is divided in various subsections dedicated to each parameter affecting the performance of the SPME technique. The webinar attendees will be guided through comprehensive understanding of the technology and the critical parameters that influence the extraction process with practical examples from already existing methods.

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