SEPTEMBER 2013 — TRENDS IN PHENOTYPIC SCREENING
Presentations on Phenotypic Screening
Single Cell Multi Parameter Analysis
Dana Nojima, Ph.D., Amgen
Watch Recorded Seminar
High content imaging can produce a large number of cellular morphometrics. Often this multiparameter data is reduced to well based averages or a single measurement per well, which can lose the richness of the data. The visualization and analysis of single cell metrics can provide a greater understanding of the data. We will illustrate this with data from cell cycle analysis with FUCCI cells and a phenotypic screen.
Phenotypic High-Throughput Assays for Hepatotoxicity and Cardiac Physiology Using Induced Pluripotent Stem Cell (iPSC)-Derived Cell Models
Evan Cromwell, Ph.D., Molecular Devices
Development of highly predictive in vitro assays is extremely important for improving the drug development process and reducing drug attrition due to toxicity reasons. iPSC-derived cell models are being adopted by the pharmaceutical industry and show great promise for toxicity evaluation and disease modeling. We have developed multi-parametric methods for toxicity assessment of pharmaceutical drugs and environmental agents. We will present phenotypic high-throughput assays using fast kinetic fluorescence, plate readers, and high content analysis measuring the impact of pharmacological compounds on cardiac physiology and general and mechanism-specific hepatotoxicity. We will describe methods for quantitative analysis of disease phenotypes and summarize results from testing focused compound libraries.
Phenotypic Screening of Cells in Solution
Michael Sjaastad, Ph.D., IntelliCyt Corporation
Watch Recorded Seminar
High content methods that provide multi-parameter measurements on single cells have revolutionized phenotypic screening. Complementary to target-based screening, phenotypic screening offers a more holistic view of drug discovery by integrating genetic, biochemical pathway and functional information into a systems view of diseases and potential therapies. The IntelliCyt iQue™ Screener utilizes a high throughput, high-content detection technology to provide information-rich data on a cell-by-cell basis. We will present several examples, including how the IntelliCyt technology has been utilized for phenotypic screening in a study assessing GFP reporters to develop Novel Acute Myeloid Leukemia (AML) therapies.
Application of Human iPSC-Derived Neurons for Phenotypic Screening and Disease Modeling
Coby Carlson, Rachel Llanas, Arne Thompson, Susan DeLaura, Eugenia Jones, and Vanessa Ott, Cellular Dynamics International
Download Presentation PDF
Millions of people suffer from neurodegenerative disorders, including Alzheimer's and Parkinson's Disease, and bettering our understanding of these debilitating conditions and finding treatments is an increasingly urgent healthcare need. A major challenge in neurological disease research and drug development, however, is access to clinically-relevant cell models. Induced pluripotent stem (iPS) cell technology offers the ability to generate such model systems. In fact, the use of human iPS cells to derive various neuronal cell types (including GABAergic and glutamatergic neurons, dopaminergic neurons, and astrocytes) is rapidly generating new opportunities for neurodegenerative disease research and drug discovery. Here we highlight the utility of iCell® Neurons (Cellular Dynamics International) in a variety of application formats in the context of a cellular system to model Alzheimer's Disease (AD). To this end, we have developed multiple endpoint assays to monitor induced neurotoxicity, measure disease-specific biomarkers, analyze epigenetic modifications of histones, and investigate the label-free response of cells to different stimuli. Together, these data illustrate how iPS cell-derived neurons can be used as a model system to study AD, and more broadly underscore the importance and potential for iPS cell technology in the future for modeling neurodegenerative diseases.
Suspension Cell Retention and Imaging Using DropArray™ Wall-less Plate Technology
Melvin Lye, Curiox Biosystems
Limited adherence of certain cell types (poorly adherent and suspension) poses a multitude of difficulties for imaging-based assays. Here, we describe the DropArrayTM a novel wall-less 96-well/384-well plate technology allowing rapid simultaneous staining of the suspension cell population in small (3µl-20µl) volumes while employing a higher-titer washing. DropArray™ offers to maintain classical throughput format of SBS-based microtiter plate while reducing the microwell volume up to 90% by using a drop format.
Using DropArray™ we conducted a time-course trafficking of five different anti-CD22 antibodies in seven B-cell lines representing four B-cell cancer types, showing shortspan antibody internalization and recycling with trafficking to lysosomes after prolonged incubations. Previously, we evaluated a bispecific antibody designed to engage cytotoxic T cells and trigger tumor cell killing, representing a functional aspect of the cancer immunotherapy.
ARI Artel Beckman Coulter BioMicroLab BioNex Solutions Biosero LLC BMG LABTECH CyBio Inc Festo Corporation Formulatrix, Inc Formulatrix, Inc Hamilton Robotics HighRes Biosolutions Hudson Robotics IntelliCyt Corporation Liconic US Western Region Molecular Devices PerkinElmer Tecan US Thermo Scientific
Senior Marketing Specialist, Sales, Brooks Account Manager, Molecular Devices Product Manager, Bio-Rad Laboratories Professor, Institute of Chemistry of Sao Carlos - University of Sao Paulo Technical Specialist, Genalyte Associate Scientist, Alios Director of Research, Amgen Inc Automation Specialist, UCSF Product Manager, ForteBio CEO, NVIGEN, Inc Scientist, Epitomics Scientist, Plexxikon General Manager, ARI , Automation Specialist, Product Specialist - Automation Solutions, Account Manager, University of California San Francisco Application Scientist, Rainin Director, Systems Engineering, Five Prime Therapeutics System Specialist, Genentech PRA, Genentech Inc. CEO, CURIOX BIOSYSTEMS Principal, Technical Representative, JL Powers & Co. Product Specialist, Agilent Technologies Applications Specialist, Artel Senior Research Associate, Roche/Genentech Regional Sales Manager, Event Specialist, Account Manager, Senior Software Engineer, Five Prime Therapeutics Sales Manager, BioMicroLab, Inc. Instructor-Coordinator, UC Berkeley Extension Scientist, consultant Sales Rep, Vice President, Senior Scientist, Technology, Genentech Senior Vice President Sales & Marketing, Microsonic Systems Consultant, Christiaanse Sales Representative , Business Unit Manager, Cellomics, Founder, Scimatic, Bioimformatics Analyst, Macrogenics Senior Software Engineer, Bio Engine Consulting Product Manager, Molecular Devices Northwest Account Manager, Director of Assay Development, Molecular Devices manager, makrocare Scientific Consultant, Application Specialist, BioMicroLab Application Scientist, PhyNexus R&D Manager,PhyNexus Sr Engineer, Amgen South San Francisco Partner, DJK Partners Account Manager, Molecular Devices Principal, North American Sales Manager, Application Scientist, Molecular Devices RSM, Account manager, FAS, PerkinElmer Product Development Engineer,
DECEMBER 2012 — ANTIBODY DISCOVERY
Combining Label Free Assay Platforms to Support Therapeutic Antibody Development from Identification of Candidate Antibodies through Pre-Clinical Development
Robin Barbour, Director of Antibody Technology, NeoTope
Label free technologies can impact antibody development from the earliest phase through entry into the clinic and beyond. Each method has its advantages and disadvantages; however by combining several of the accepted technologies a company can leverage the strengths of each. The plate based label free instruments can be incorporated into the identification of antibodies that block cell adhesion, or GPCR signaling and are useful in a high throughput screen for antigen specific clones. The Forte-bio Octet allows for rapid ranking of positive clones, a quick and easy method to monitor antibody production and a method to quickly determine relative affinities during engineering. The BIAcore T200 offers the gold standard in kinetic evaluations, and provides a rich data source for antibody characterization.
New Technologies enhancing hybridoma production & development: The Implementation of Genetix ClonePix FL
Monica You, Associate Manager, Elan Pharmaceuticals
Watch Recorded Seminar
Biologics: Special Considerations for Toxicology Testing
Meredith Rocca, Janssen Pharmaceuticals
Watch Recorded Seminar
Biologics pose unique challenges for nonclinical toxicology testing. Although the general principles of nonclinical toxicity testing are the same for biologics and small molecule drugs, additional factors to consider include species relevance, long half-life, the potential for immunogenicity, and the possibility of performing the FIH study in a patient population. Successful strategies for dealing with these issues will be discussed.
Label-Free Binding Assays: What Lies Beneath the Curve
Pui Seto, Scientists, Pall ForteBio
Download Presentation PDF
The rapid growth over the last few years of antibody and protein therapeutics has created a need for fast, reliable and versatile screening tools to engineer improvements in biological drug candidates. Kinetic analysis of binding of target molecules to antibody and other protein drug candidates is critical to the selection of clones for development and for testing functional binding characteristics throughout development and production. Direct binding, real-time assays on the Octet and BLItz systems provide ease-of-use, fast assay development, enhanced throughput, versatility in assay design and the ability to rapidly screen molecular libraries to accelerate the discovery process.
AssayMAP- A Quantitative, Automated, Analytical Sample Preparation Platform for Biologics Characterization
Mitu Chaudhury, Product Manager, Sample Preparation Solutions, Agilent Technologies, Inc.
Download Presentation PDF
The FDA’s QbD initiative and the rise of biosimilars are placing tremendous strain on biopharmaceutical process development and quality control groups. Protein analysis methods must be developed that deliver higher precision, sensitivity and throughput, in order to meet these new demands. A wide range of analytical instruments (such as CE, HPLC, LC-MS) is used to examine structural characteristics of proteins such as posttranslational modification, but the protein of interest must first be highly purified intact from complex sample matrices (such as culture supernatant or plasma) and often requires other complex preparation methods (such as enzymatic treatment or labeling). Overall workflows can involve highly complex combinations of sample processing methods. Several techniques for sensitive and precise protein quantitation are widely used, including affinity HPLC and immunoassays, but these methods suffer from low throughput and complexity of automation, respectively.
Here we present a new quantitative, high throughput sample preparation platform based on disposable cartridges operated on the AssayMAP Bravo system — a modified 96-channel, microplate-compatible liquid handler equipped with ultra-low dead volume syringes that connect to the cartridges through a special probe. The liquid handler provides highly precise positive-displacement flow control through the cartridges in either direction to well under 1 µL/min, and can dispense liquids through the cartridges at 20 bar pressure. Applications presented will include:
- Affinity chromatography for quantitation and further analysis
- Peptide mapping workflows- purification, digestion, clean-up and desalting
- Complete, automated sample preparation for N-glycan analysis
OCTOBER 2011 — ADVANCES IN CELL-BASED SCREENING ASSAYS
- Susan Watson, Xoma Pharmaceuticals
High Throughput Flow Cytometry for Screening Antibodies Produced from Phage Library
- László G. Kömüves, Genentech Inc.
Digital Pathology and BioImage Informatics: Quantitative Microscopy on a Large Scale for Drug Discovery and Development
- Matt Clutter, Stanford University
Automated Phospho-specific Flow Cytometry for Screening Endogenous Signaling Pathways in Primary Blood Cells
- Anthony Pitt, Lonza Cologne GmBH
High-content Analysis of Signaling Networks Using Protein Fragment Complementation Assays (PCA)
- Brad Larson, BioTek Instruments
Automated, Hepatocyte-Based Methods in Early In-Vitro ADME/Tox Testing: The Importance of Cell Model
- Technology Forum featuring short presentations from:
Curiox Biosystems Pte Ltd, Horizon Discovery Ltd., Tecan, and LabCyte Corp.
JUNE 2011 —THE NEXT GENERATION OF IMMUNOASSAYS: TRENDS IN ELISA INNOVATION AND AUTOMATION
- Zhao Ren, Elan Pharmaceuticals
AlphaScreen Technology: A Robust and Versatile Platform for Assay Development and HTS
- Brett Maters, Bioscale
BioScale ViBE: A Versatile High-sensitivity Solution, Improving Efficiency, and Reducing Total Time to Quality Results
- Rob Durham, Gyros
Evaluation of Tau Phosphorylation Using a Non-traditional Immunoassay Platform
- Ruth Motter, Elan Pharmaceuticals
Beyond Liquid Handling — Using the Hamilton Microlab STAR to Increase Immunoassay Efficiency
- Sheila Keating, Blood Systems Research Institute Laboratory
Frenemies: Using Multiplexed Immunoassays to Study Immune Responses to Infectious Diseases
- New Technology Forum featuring short presentations from:
Tecan, Hamilton, CisBio, Cell Biosciences, Curiox and LIMR Chemical Genomics Center
JUNE 2010 — ADVANCES IN LABEL-FREE DETECTION AND ANALYSIS
- John C. Owicki, Ph.D., Owicki Consulting
Surface attachment in label-free assays: effects on binding affinity and kinetics
- Robin Barbour, Director, Neotope
Integrating label-free technologies into your drug discovery workflow
- Yasmina Abdiche, Ph.D. Senior Principal Scientist, Rinat-Pfizer
Use of disposable label-free real-time biosensors in epitope binning monoclonal antibodies
- Tony Giannetti, Ph.D. Research Scientist, Genentech
Integration of SPR into small molecule drug discovery
- Michael Bova, Ph.D., Director of Target Advancement, Élan
Identification of compounds that inhibit cell adhesion using BIND label-free technology
- Emmanuel Sturchler, Ph.D. , Scripps Research Institute, Scripps Florida
Characterization of GABAB receptor allosteric modulators using the CellKey® System
DECEMBER 2009 — INTEGRATING NEW AUTOMATION TOOLS INTO THE LABORATORY WORKFLOW
Introduction & Opening Remarks
Mike Biros, Amer El-Hage, LRIG Bay Area.
Accelerating Research by Integrating Vision and Motion Measurement Technologies into the Laboratory Workflow
Christoph Wimmer, Business Development Manager, National Instrument
By integrating measurements and automating processes, labs can accelerate research and simplify laboratory processes. However, determining which tools to use to automate labs is a challenge that many researchers currently face. Using vision and motion technologies, researchers can control and automate instruments with flexibility, easily integrate with off-the-shelf hardware, and quickly and accurately solve diverse application challenges from automating test equipment and research labs to controlling biomedical sampling and testing. Explore how to deploy each of these technologies into lab environments to accelerate research projects by increasing performance, reliability, and productivity. Also review customer solutions that showcase how companies like Celera Genomics Group automated pipetting robots that transfer samples and dispense reagents while processing initial E. coli clones; how Wyeth Neuroscience used computer-based data acquisition and NI LabVIEW to automate the generation of synaptic physiology data from multiple brain slices; and how SpectruMedix automated a commercial DNA-sequencing capillary electrophoresis spectrometer.
A 4-Way Nano Fluidic Intersection: When Scientists and Engineers Collide
Christopher A. Bonagura, Ph.D., Art Robbins
ARI has developed and released a new nano fluidic liquid handler that accompanies the Phoenix platform. This new system allows for both air backed and liquid backed fluid handling capabilities, depending on the user’s needs. The nano tip dispenses down to 100 nl with high precision. The system now comes configured as a single channel unit and up to a 4-Way “Quad” channel unit, combined with a 96 or 384 channel head of the Phoenix. The 4-channel allows for lab personnel to independently use each of the 4 dispense channels to be tasked to separate activities, allowing for new combinations of complex reactions and additions to be done quickly and accurately.
We will describe developing the new ARI nano dispenser technology and the software that drives it. We illustrate how features of the system allow users in protein crystallization to generate novel methods that could not be achieved using other less versatile systems. The Phoenix can also be fitted onto larger integrated robots as demonstrated by the SE unit from Rigaku, and an earlier integrated fluid handling system, the CrystalBot. Finally, we will show how this technology can be deployed as an advantage in other high throughput laboratory settings.
Overlord2 - Access to a Huge Range of Instruments from Your Existing Laboratory Automation Application
Malcolm Crook, Ph.D. & Rob Harkness, Process Analysis & Automation
New hardware and software ideas are required to advance existing laboratory automation workcells into smaller, reliable, easier to use and flexible sample processing units. This can be achieved by the clever integration of technologies from other factory automation systems and the clever design of control software. An example of robust engineering, new systems integration control software and a new user front end will be presented as new tools for a new decade of science.
Vendor Exhibits and Networking
Ultrasonic Microplate Mixing for Efficient Sample Prep and Improved Assay Results in the Laboratory workflow
Vibhu Vivek, Founder and CTO, Microsonics
The issues of proper assay mixing, compound re-solubilization, isothermal thawing and bead and cell suspension have become more acute as microplate volumes have decreased.
Mixing has proven to be a limiting factor in the laboratory automation industry’s migration to smaller well volumes and higher density plate formats for compound and assay microplates. As the plates move to higher densities, the aspect ratio of the wells, the surface tension of the fluid and capillary forces all make conventional mixing a more difficult task.
Microsonics has harnessed the power of ultrasonic energy for effective non-contact mixing, solubilization, isothermal thawing and bead and particle re-suspension in microplate format.
Its patented technology uses a unique transducer to produce strong Lateral Ultrasonic Thrust TM (LUT) waves in liquids, creating very effective non-contact mixing and thawing for 96, 384 and 1536 well microplates in the form of a commercially available product, the HENDRIX SM100. This novel, patented form of ultrasonic energy is generated using Fresnel Annular Sector Actuator-based (FASA) transducer elements and introduced into microplate wells with finely tuned control of the amount and type of power, and less dependence on plate material and geometry. This allows everything from gentle mixing of cells, to more aggressive mixing of assay components, magnetic beads and particles to more energetic thawing of frozen microplates or mini-tubes, all without any contact to the material in the microplate wells!
Numerous examples will be cited, as well as videos dramatically illustrating the unique properties of this outstanding new technology now commercially available as the HENDRIX SM100 for effective fluid processing.
Direct Drive Technology for Laboratory Automation
Marc Valer, Integrated Systems Product Manager, Agilent – Velocity 11
With the application of direct drive technology and extensive input from life science automation scientists, Agilent designed the Direct Drive Robot (DDR) from scratch. It’s a full size robot that, when integrated with VWorks scheduling software, effectively handles microplates with more flexibility, better efficiency and improved safety compared to its predecessors. The integration of direct drive technology and advanced collision sensing among other novel features, makes the DDR the first robot that can be truly utilized in a top performing encapsulated solution as well as a benchtop, mobile system that effectively solves integration needs where they arise, while at the same time defining a new level of protection for equipment and operators in life sciences.
We will present some of the new hardware and software concepts and algorithms utilized for advanced path planning, re-gripping and collision detection/recovery as well as innovative solutions needed for robust manufacturing and testing.
An Integrated, Automated System for High-Throughput ADME Sample analysis to Enhance Laboratory Workflows
Michael Biros, Biocius (A wholly owned subsidiary of BioTrove)
Efficient and robust in vitro ADME assay screening is essential for early drug candidate optimization. Mass spectrometry (MS) has emerged as the preferred method to address the multi-dimensional challenge toward compound selection on account of its sensitivity, selectivity, and label-free rationale. However, a daunting shortcoming of MS (in contrast with competing technologies that utilize fluorometric or radiometric probes) is its low throughput: Traditional MS approaches require time-consuming off-line sample preparation and liquid chromatography (LC) separation. In contrast, the innovative RapidFire 300 instrument draws on MS-based data quality without conceding speed and integrates easily into existing laboratory workflows.
At its core, the RapidFire 300 platform consists of a fast, serial, solid-phase extraction (SPE) sample purification system, interfaced to an atmospheric phase ionization (API) MS. Automated sampling of analytes is achieved using a plate handler with a barcode scanner that can accommodate 96-well or 384-well plate formats. Fluidic flow is monitored by an optical sensor to detect aspiration of each sample. Samples are aspirated and delivered to an on-line SPE cartridge for purification. The SPE cartridge cassette can hold up to six different cartridges, allowing automated cartridge changes or multiple packing chemistries for method development applications. Analytes of interest are retained on the cartridge, washed, and eluted. The eluted analytes are delivered to a triple quadrupole (QqQ) or time-of-flight (TOF) MS for ionization and mass detection. A single sample can be purified, delivered, and detected by the mass spectrometer in 6-10 seconds and complete cycling of a 384-well plate can thus be achieved in less than an hour.
This novel automated SPE-MS technology integrates into the work flow of standard HTS laboratories streamlining and enabling an array of HTS and in vitro ADME studies. Thus, the RapidFire platform provides an innovative solution for label-free screening in high-throughput drug discovery and development applications.
Concluding Remarks and Event Prizes
JUNE 2008 — TRENDS IN FLUORESCENCE-BASED IMAGAING ASSAYS
Characterization of Cytotoxicity in Neuronal Cells using the Cellomics Neuronal Profiling Bioapplication
Michael Bova, Ph.D., Elan Pharmaeceuticals
Easy Exploration of High-Content Data with AcuityXpress
Pierre Turpin, Ph.D., MDS/Molecular Devices Corporation
Advances in Higher-Throughput Live Cell Assays under Laminar Flow
Carolyn G. Conant, Ph.D., Fluxion Biosciences
High-Throughput Imaging of Zebrafish Embryos for Drug Discovery
Steve Miller, Ph.D., Blueshift Biotechnologies
MAY 2007 — TRENDS IN ACCELERATING DRUG DISCOVERY RESEARCH
Parallel Drug Lead Optimization with Early ADMET
John Wang, Ph.D., Associate Director, Novartis
An Integrated Approach to Process Automation in Research and Development at Medarex, Inc.
Denise Williams, Assistant Director, Research Automation Services, Medarex, Inc.
Trends in Sample Storage Automation
W. Steven Fillers, Ph.D., VP & CSO, TekCel LLC
A Simple and Rapid Methodology for Determination of Enzyme/Substrate Specificity using Micro Parallel Liquid Chromatography
Jeff Koehler, Principle Scientist, Nanostream
BioTek’s Synergy™ 2, A Multi-Detection Microplate Reader Capable of Quantitating cAMP and TNF-a Using Various Homogeneous Assay HTS Technologies
Paul Held, Ph.D., Senior Scientist, BioTek Instruments
High Throughput Screening via RapidFire™ Mass Spectrometry
Can Ozbal, BioTrove, Inc.
The Latest in Screening Data Management with ActivityBase XE
Sermed Naaman, IDBS
APRIL 2006 — ADVANCES IN FLUORESCENCE-BASED CELLULAR ASSAYS
Mike Biros, MBA, LRIG Bay Area
DiscoveRx PathHunter Assays: Non-Imaging Translocation Detection For Pathway Profiling
Keith Olson, Ph.D., DiscoveRx
HaloTag: A Multipurpose Fusion Protein Tag for Analysis of Dynamic Cellular Events
Randall Learish, Ph.D., Promega
Novel Automated Imaging Approach for Multi-parametric Measurements of Cytotoxicity
Michael Honeysett, MBA, GE Healthcare
Molecular Devices Total Imaging Solution Case Study: ImageXpress Ultra Confocal, Transfluor and AcuityXpress Informatics Analysis
Pierre Turpin, Ph.D., Molecular Devices Corporation
EasyCyte Volumetric Flow Cytometer & GuavaLink Automation Software: A flexible System to Automate Fluorescence Screening of In Vitro Toxicology, Cell Counting, Cell Based and IgG Quantitation Assays
Ben Alderete, Guava Technologies
PerkinElmer: New Developments in Homogenous AlphaScreen and LANCE Assays
Allan Atkinson Ph.D.
MARCH 2005 — CURRENT BOTTLENECKS IN LABORATORY AUTOMATION
Are you compatible with today's automated laboratory?
Mike Biros, LRIG Bay Area Executive Chair, Mountain View
Building a fully integrated screening operation from scratch
Ge Wu, Ph.D., Director Assay Development and Automation, FivePrime, South San Francisco
Setting up an automated high-throughput screening lab in academia
David E. Solow-Cordero, Ph.D., Associate Director, High-Throughput Bioscience Center, Department of Molecular Pharmacology, Stanford University School of Medicine
Bottlenecks in lab automation: constraints on sequencing automation
Steven Wilson, Instrumentation Specialist, DOE Joint Genome Initiate, Lawrence Berkeley Laboratories, Walnut Creek
NOVEMBER 2004 — ADVANCES IN FLUORESCENCE ASSAY TECHNOLOGIES
Implementing intra-well assay controls using the CellCard™ System
Ion channel assay development on the synergy HT multi-detection microplate reader from Bio-Tek
Quantitation of cell health in high content screening assays
Two-dimensional anisotropy for homogenous object-based fluorimetry
Using flow cytometry to enhance signal:noise in cell-based assays
Use of microcytometry in cell health monitoring and secondary screening applications
FEBRUARY 2004 — VALIDATION OF LABORATORY LIQUID HANDLING SYSTEMS
SEPTEMBER 2003 — OPEN COMMUNICATION STANDARDS AND APPROACHES FOR LABORATORY AUTOMATION
APRIL 2004 — HTS: RECENT ADVANCES IN AUTOMATING KINASE ASSAYS