Biacore System description. GE Healthcare. Biacore label-free interaction analysis. Optional software packages
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- Egbert Adam Blankenship
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1 GE Healthcare Biacore label-free interaction analysis Biacore 4000 Biacore 4000 offers a complete solution for large-scale, label-free molecular interaction analysis, delivering high throughput without compromising on data quality (Fig 1). The system hardware and software provide innovative solutions for maximizing productivity in key areas such as antibody selection, biotherapeutic and small molecule (LMW) drug discovery. Key benefits: High sensitivity and high quality interaction data for better-informed decision making 60 h unattended operation with parallel analysis of up to 16 targets Large-scale molecular analysis generating up to 4800 interactions in 24 h Seamless LIMS integration with convenient data import and export functionality Innovative software solutions for increased productivity and performance System description Biacore 4000 delivers high quality kinetic, affinity, concentration, specificity, and binding data in screening assays and detailed characterization studies. The system is designed for large-scale, parallel interaction analysis and can be used during multiple phases of drug discovery, from early screening to characterization. Fig 1. Biacore 4000 is a molecular interaction analysis system for rapid screening and characterization. Optional software packages Pharmaceutical applications, including LMW drug discovery and biotherapeutics, face unique challenges and demands. GE Healthcare offers add-on software packages with dedicated tools optimized for two key areas. Biacore 4000 LMW Extension Package: Optimized for small molecule drug discovery, especially fragment screening. Biacore 4000 Antibody Extension Package: Optimized for screening and characterization of antibodies and antibody fragments. Main applications: Fragment and LMW compound screening Compound hit validation, hit-to-lead characterization, and lead optimization of compounds Antibody and antibody fragment screening and characterization Data file, AD
2 System features Biacore 4000 has a flexible format providing possibilities for parallel multiplexed assays and efficient assay development. Microfluidics optimized for throughput and flexibility The innovative design of the flow system increases sample throughput and allows several different experimental conditions to be tested in parallel, accelerating assay development. The system contains four independent flow cells, each with its own needle, enabling parallel injections. Different interactants can be immobilized on the five different detection spots in each flow cell, making different assay configurations possible. Figure 2 shows two principal types of parallel multiplexing, optimized for different experimental objectives. In an assay configuration optimized for throughput (Fig 2A), four different interactants are immobilized on each flow cell and four different samples are analyzed in parallel. In Figure 2B, a configuration maximizing the information obtained per sample is shown, where one sample is assayed with 16 different immobilized interactants simultaneously. In both assay setups, 16 interactions can be detected per cycle or up to 4800 interactions in 24 h. (A) Optimized for sample throughput 4 4 immobilized interactants = 16 interactions/cycle 4 samples Different samples in each flow cell (B) Optimized for information per sample 16 immobilized interactants = 16 interactions/cycle 1 sample Same sample in all flow cells Reference spot Reference spot Reference spot Reference spot Reference spot Reference spot Reference spot Reference spot Fig 2. Two principal assay configurations are shown. (A) Up to 1200 samples with four different immobilized interactants can be run in 24 h. (B) Up to 300 samples with 16 different immobilized interactants can be run in 24 h. Fig 3. Biacore 4000 supports both 96- and 384-well microplates and up to 10 rack trays can be loaded and run unattended. System hardware designed for productivity Biacore 4000 can perform 60 h of unattended operation, reducing labor and increasing productivity. In the rack hotel, up to 10 rack trays (microplates and reagent rack) can be loaded with automatic rack exchange. The system supports both 96- and 384-well microplates (Fig 3). Temperature in the system is stringently controlled (4 C to 40 C) allowing interactions to be accurately monitored at biologically relevant temperatures. In addition, the sample compartment is temperature controlled, allowing samples to be kept warm to aid solubility or sensitive samples to be kept cool. Solutions for workflow integration Biacore 4000 software includes solutions for system integration with Laboratory Information Management Systems (LIMS) to simplify import and export of sample data, run information, and results. The sample manager imports data from XML, text, or Microsoft Excel files. Data can also be retrieved from a database using Web Service, a tool that provides direct access to Biacore 4000 database and enables automated transfer of data to and from LIMS. A barcode scanner is included in the system for tracking microplates AD
3 Database solution tailored for large-scale applications To provide the storage capacity required by large-scale assays, all data is stored in an Oracle database located on a server. This database setup provides searchable data and security features inherent to Oracle database systems. Data can be accessed from separate workstations, allowing users to control runs and evaluate data from different computers. Monitor interactions in real-time As the assay progresses, data are shown in real-time. The data can be viewed in different formats, such as sensorgrams from each flow cell (Fig 5). Note: The Oracle client is provided with the system; however, the server and Oracle database management software are not included. Intuitive software for rapid data processing and analysis Biacore 4000 software maximizes productivity, while maintaining flexibility and user friendliness. Quickly create methods Biacore 4000 Control Software provides full flexibility to design methods for a wide range of assays. Run methods are setup in logical steps, and for added efficiency, predefined methods are available that can be used directly or modified to suit the user s needs. The control software has an intuitive graphical interface for simple viewing and editing of the method (Fig 4). Fig 5. Real-time data displayed as the assay progresses. Evaluate large data sets rapidly Biacore 4000 Evaluation Software utilizes several automated tools to speed up analysis of large data sets, including automated quality control tools to identify infrequent sensorgram disturbances, predefined evaluation methods, and sophisticated validation algorithms for kinetics and affinity. In the evaluation software workflow, the selected data is inspected and approved prior to evaluation (Fig 6). Select data Approve data Spot settings, select curves, signal correction, quality control Evaluate data Kinetics and affinity, screening, concentration, assay development Fig 4. The control software gives a comprehensive overview of the run method before the run (upper panel) and at the start of the run (lower panel). Fig 6. Evaluation software workflow AD 3
4 Automatic QC assessment for rapid approval of large data sets The quality control (QC) assessment tool allows thousands of sensorgrams in a data set to be rapidly checked for approval. This tool uses advanced algorithms to automatically identify deviants, and gives the user full control to decide which data to exclude (Fig 7). Data Set Application-tailored evaluation tools for efficient data processing Biacore 4000 Evaluation Software provides applicationtailored evaluation support to facilitate kinetics, affinity, and concentration analyses. For convenience, an overview of the data is shown as thumbnails combined with details of selected data (Fig 9). Evaluation is streamlined, requiring only a small number of clearly defined operations to go from approved data set to final results. <acceptable?> Recalculation No Yes Approve Exclude Evaluation Fig 7. Using the QC assessment tool, large data sets are quickly examined after a few iterations using the workflow above. To identify deviants, the QC assessment tool plots sensorgram data based on QC descriptors and maps any deviation from the representative mean (Fig 8). QC descriptors identify infrequent sensorgram disturbances and enable the user to exclude deviating sensorgrams. This valuable tool detects all major and > 95% of minor deviations, and allows large data sets of several thousand sensorgrams to be inspected in less than an hour. In addition, trendplots can be used to identify carry-over between cycles or binding to reference, for example. After a few iterations, the data is ready for evaluation. Fig 9. A complete overview of the data is given using thumbnails (to the left) with a detailed view of selected data (to the right). Decreased time-to-results with powerful plot tools for screening and ranking Biacore 4000 Evaluation Software offers a range of dedicated plot tools that automate evaluation, saving days of work. In the evaluation software, report point values can be plotted in a number of ways for different evaluation purposes. Ranking borders and cut-off lines can be added to plots of single report points against variable values. Data in plots can be tagged using user-defined curve markers. The marker information is automatically retained in subsequent plots, result tables, or further processed data for simplified data evaluation (Fig 10). Fig 8. A QC descriptor plot, to the left, shows deviations from a representative mean. The sensorgram for a selected deviant is displayed in the right panel for closer inspection, so that the user can decide whether or not to exclude it. Fig 10. Data can be tagged using user-defined curve markers (marked in red). In this plot, ranking borders classify high, medium, and low level binders AD
5 Sophisticated tools for evaluation of kinetics and affinity Biacore 4000 Evaluation Software provides streamlined functions for evaluation of kinetic and affinity data. Multiple ligand-analyte pairs can be evaluated in parallel if they are analyzed in the same run. Evaluation is rapid, and the software detects and recommends exclusion of sensorgrams that are not appropriate for the analysis (Fig 11). The software uses a 1:1 binding algorithm to derive kinetic and affinity constants (Fig 12). To aid data interpretation, the quality of the fitting results is validated against a number of rules defined by sophisticated algorithms. The validation rules take into account, for example, closeness-of-fitting and significance of derived parameters. Dedicated tools for reliable concentration measurements Evaluation tools for concentration analysis provide automatic generation of calibration curves and calculation of sample concentrations (Fig 13). The coefficient of variation (CV) for replicate samples is also provided automatically. Fig 11. Data validation tools use advanced algorithms to check data quality. In the example above, three data sets (marked with an X) are suggested to be excluded from further evaluation. Fig 13. Concentration analysis tools automatically generate calibration data, and calculate sample concentration and variation between replicates. Fig 12. Kinetic and affinity constants are derived by the software using a 1:1 binding algorithm. Fitted curves are superimposed in each concentration series AD 5
6 Seamless transition into regulated environments Biacore 4000 software facilitates compliance with worldwide regulatory expectations by offering Validation Services and 21 CFR Part 11 functionality. The software is developed in accordance with an accepted development model to ensure adequate validation and includes 21 CFR Part 11 functionality such as: User authorization levels: administrator and user levels set access rights to software functions Published procedures for operational control: enables run and evaluation settings to be locked together in routine assays Audit trails: tracks record modifications and maintains complete version histories for published procedures Electronic signatures: for published procedures Data integrity: access control and enforced version handling Validation support documentation such as System Assessment Report, Software Conformance and Hardware Conformance Certificates is provided. Validation services GE Healthcare Validation GxP Services provide a comprehensive suite of validation services to support equipment through its entire lifecycle, including retirement and decommissioning (Fig 14). Services include GxP impact assessments, installation and operational qualifications, change controls, and requalifications. System qualification is performed by certified, GxP trained service engineers and uses a risk-based approach in alignment with recent guidelines such as GAMP (Good Automated Manufacturing Practice) and ASTM (American Society for Testing and Materials). Application support for increased productivity GE Healthcare offers two add-on software packages with dedicated tools optimized for rapid screening and characterization of small molecules and antibodies. Biacore 4000 LMW Extension Package Biacore 4000 LMW Extension Package provides a powerful solution for efficient and confident selection of LMW compounds including fragments, without compromising on either data quality or throughput. Tailored software tools speed up assay setup and enable streamlined evaluation. Rapidly clean your library from sticky substances that may otherwise interfere with assay performance Confidently discriminate well-behaved binders from promiscuous or atypical binders Analyze affinities into the mm range for LMW compounds and fragments before scaling up target production Conveniently export data to analysis software such as Spotfire or Microsoft Excel for customized evaluation Characterize a wider range of compound-target interactions using additional kinetic fitting models Biacore 4000 Antibody Extension Package Biacore 4000 Antibody Extension Package maximizes productivity in large-scale antibody analyses, saving both time and reagents during antibody selection and biotherapeutic development. This software solution increases throughput and shortens time-to-result. Save time and reagents during antibody screening and characterization - more efficient assay setup using additional spot-addressing functionality Obtain valuable kinetic information in half the time using small volumes of crude samples with 2-over-2 kinetics Save days in analysis time with dedicated software for pairwise epitope mapping Simplify customized data analysis using additional functionality for export to analysis software Address a broader range of antibody-antigen interactions using additional kinetic models Fig 14. GE Healthcare Validation GxP Services provide comprehensive support thoughout the equipment s entire lifecycle AD
7 Biacore 4000 specifications Performance characteristics Typical throughput Typical data evaluation time: Up to 4800 interactions/24 h (direct binding assay, 4 targets per flow cell, cycle time 5 min) LMW compound screening: 1200 samples/24 h Antibody screening: 960 samples/24 h Kinetic characterization: 240 determinations/24 h (4 targets per flow cell) Screening: < 1 h Kinetics: 1 to 2 h Technical specifications and characteristics Detection technology Surface plasmon resonance (SPR) biosensor Working principle Parallel processing of four independent flow cells, each containing five detection spots All spots addressable, but typically used as addressable + 1 reference Type of information Binding kinetic, affinity, selectivity, and specificity data. Concentration measurements Sample type LMW drug candidates to high molecular weight proteins in various sample environments, e.g., in DMSO-containing buffers, cell culture supernatants, or serum Data collection rate 1 Hz (default) or 10 Hz Kinetics Association rate constant (k a ): Dissociation rate constant (k d ): 10 3 to 10 9 M -1 s -1 (LMW compounds: M -1 s -1 ) to 1 s -1 Detection limits 100 pm Molecular weight detection Analytes with relative molecular mass (M r ) > 50 (Da) Immobilized interactant consumption Typically 3 to 10 µg per immobilized spot (varies with application and size of immobilized molecule) Sample consumption Typically 60 µl per flow cell (range 30 to 425 µl) Rack hotel capacity 10 rack trays, each with one microplate (384- or 96-well) and one 24-well reagent plate Typical sample capacity per run Antibody screening: well microplates LMW compound screening: well microplates Kinetic characterization: well microplates Automation 60 h unattended operation Sample temperature 4 C to 40 C (maximum cooling 18 C below ambient) Analysis temperature 4 C to 40 C (maximum cooling 20 C below ambient) Baseline noise < 0.1 RU (RMS) Baseline drift Typically < 0.3 RU/min in ambient temperature range 15 C to 35 C In-line reference subtraction Automatic, if selected In-line buffer degassing Automatic Dimensions (W H D) Instrument: mm Instrument on trolley: 1800 ( ) 623 mm Net weight Instrument 188 kg Trolley 82 kg Thermostat unit (including tubing and coolant) 36 kg Mains requirements Processing unit and system controller Autorange VAC, Hz, protective earthing Thermostat unit 230 VAC/50 or 60 Hz 115 VAC/60 Hz 100 VAC/50-60 Hz Power consumption Processing unit Maximum 6.3 A (at 100 VAC) System controller Maximum 7.2 A (at 100 VAC) Thermostat unit Maximum 12 A (at 230 VAC) Maximum 11 A (at 115 and 100 VAC) Data handling and storage PC operating system Compatible with Microsoft Windows TM XP (32-bit ), and Windows 7 (32-bit and 64-bit), and Windows 8.1 (64-bit) Database storage Compatible with Oracle , Oracle , Oracle , Oracle , and Oracle Interfacing Web Service for automatic import of sample data and export of results from and to LIMS Export formats: XML and tab delimited text Import formats: Excel, XML, and tab delimited text Compliance Safety and EMC standards GxP Complies with and applies to Europe and North America (US and Can) standards Validated software with 21 CFR Part 11 (ER/ES) functionality Delivery: Biacore 4000 includes a processing unit, control and evaluation software, Windows operating system, barcode scanner, and a dedicated trolley. Computer, external thermostat unit, and desktop package must be ordered using separate order codes. Optional: Validation support documentation and GxP services for facilitated regulatory compliance, Biacore 4000 LMW Extension Package, and Biacore 4000 Antibody Extension Package. On-site requirements: Database server with Oracle , Oracle , Oracle , Oracle , or Oracle installed. Contact your local GE Healthcare representative for the latest information regarding on-site requirements AD 7
8 Ordering information System * Product code Biacore * Includes a processing unit, control and evaluation software, barcode scanner, and a dedicated trolley. Computer, external thermostat unit, and desktop package must be ordered on separate order codes. Optional packages Product code Biacore 4000 LMW Extension Package Biacore 4000 Antibody Extension Package GE Healthcare Validation GxP Services For more information, please contact your local GE Healthcare representative. Related literature Product code Biacore 4000 LMW Extension Package, Data file Biacore 4000 Antibody Extension Package, Data file GE Healthcare UK Limited Amersham Place Little Chalfont Buckinghamshire, HP7 9NA UK GE, GE monogram, and Biacore are trademarks of General Electric Company. Excel and Windows are trademarks of Microsoft Inc. Oracle is a trademark of Oracle Corporation, Spotfire is a trademark of TIBCO Software Inc. All other trademarks are the property of their respective owner General Electric Company. First published Jan All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. A copy of these terms and conditions is available on request. Contact your local GE Healthcare representative for the most current information. GE Healthcare Bio-Sciences AB, Björkgatan 30, Uppsala, Sweden GE Healthcare Europe, GmbH, Munzinger Strasse 5, D Freiburg, Germany GE Healthcare Bio-Sciences Corp., 800 Centennial Avenue, P.O. Box 1327, Piscataway, NJ , USA GE Healthcare Japan Corporation, Sanken Bldg., , Hyakunincho, Shinjuku-ku, Tokyo , Japan For local office contact information, visit AD 10/2015
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