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The HPGETM 1000 System from LabIntelligence, Belmont, California, is the first fully automated, integrated gel electrophoresis system with real-time fluorescence detection, incorporating size selective concentration, separation and zone selective retrieval via automated on-line electroelution.
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The system's spectrophotometric abilities allow for sensitive multi-color detection of peptides, proteins, carbohydrates, nucleic acids, chromosomes, even cell particles and viruses in routine and research. Its Windows'95 AI-assisted, GLP/GMP compliant user interface helps scientists in demand for rapid development of new quantitative, analytical and preparative applications in the life sciences.
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The Idea
An objective design consideration for an integrated electrophoretic system had to include the following aspects: a dynamic control with active feedback loop for all parameters involved in the separation process to ensure accurate and reproducible experiments; the ability to pre-concentrate sample components of low concentration or for in-situ sample cleanup prior to the separation process; a capability to monitor the separation progress dynamically, preferrably via photometry; a capacity to perform on-gel assays for direct determination of the biological activity; the ability to elute or retrieve separated components selectively for immobilization, preperative use, other post-separation processes or multidimensional analysis; and a concept for information management capable to store, process, compare and evalute experimental data in strict compliance with all guidelines for GLP (Good Laboratory Practice) for forensic and certified laboratories. Furthermore, the hardware had to be capable of different gel types, and the software had to fullfill all criteria that apply for a user friendly operation interface and representative data presentation.
Why HPGE?
- Fully automated real-time analysis system - Automatical sample application under electrical fieldstrength applied [=better starting zone] - Selective concentration of components possible [e.g. virus enrichment independent of the content of components with low molecular weight] - Feedback regulation, monitoring and control of all parameters during all phases of the experiment [e.g.. buffer capacity, fieldstrength within the separation medium, total voltage, current, temperature...] - no post-treatment of the gel after termination of the separation required for zone visualization [Real time detection]- immediate information on separation dynamics [direct detection of mobility shifts during the separation or of irregluar molecule movement, e.g. reptation or conformational changes can be determined] - Ability to perform direct In-GelTM Assays to determine biological activities with built-in capability to incubate and photometric kinetics measurement [e.g.. for enzyme activity or antibody activity determination]
- Fluorecence photometer with high sensitivity [sub-nanogram level, dependent on labeling reagents] and linearity [4 orders of magnitude dynamic range] - Broad spectral detection range allows for use of different fluorescence labels, utilizing tag emission maxima within the entire visible range - Fluorescence spectrophotometer for multi-color labeling protocols [HPGE 1000S only] - Built-in pulse field/field inversion power supply - Built-in "Hold"-mode to avoid zone diffusion - Built-in potential for temperature gradient electrophoresis [TGE] [Cooling/Heating range 0°C...+70°C, temperature ramping over time] - Gel format and transparent plastic gel trays provide for use of semi-liquid separation media for separation of viruses, cells, cell particles and chromosomes - Automatic sample cleanup or Selective ConcentrationTM prior to the separation process ensures clean separations with low background - Selective Zone RetrievalTM: a quantitative advantage over blotting, with immediate control of transfer efficiency via pre/post-scanning function at the position of the electro-elution - In-GelTM Assay: direct estimation of biological activity or functionality of separated components via enzyme-substrate or antigen-antibody kinetics in the gel itself - Complete physical characterization of molecules [Ferguson-Plot-Analysis], using gels of different gel concentrations within a single experiment - GLP-compliant data for forensic and certified laboratories: · User hierarchy and level-based privilegies · Report function for printout and on-screen display with information on all instrumental parameters during the entire time of the experiment · Data validation management [LIMS-function]· Standardized data base with import/export-capability and data portability for DOS or spreadsheet applications · Sample comparison and pattern recognition - Instrument's small footprint allows operation also in laboratories with limited bench space
- Operating software fully integrated Windows3.x®/'95®/'98®/NT® application: integrated graphical user interface with AI [Artificial Intelligence] support, high resolution graphics [with color printout on Inkjet-printer] and support for data import/export - Self explainatory icons and instrument controls [check for parameter plausibility and logics support] - Only few manual user interactions necessary [guidance-positioned buffer containers, gel trays, applicators] - Pre-defined routine methods for protein, carbohydrate and DNA analysis - Simple configuration procedure for method parameters with user guidance during all experimental phases - Additional evaluation software for special separation problems or enhanced evaluations - CurveFit: general curve-fitting program for non-linear calibrations - MOLWT: molecular weight determination [in Daltons/Base pairs] - LPATHTM: elongated gel evaluation for wide-range DNA-separations [50bp-2Mbp] via LongPathTM Technique - TCALCTM: separation predictions based on extrapolation obtained from separation histories [dynamic mobility and resolution evaluation]
- Complete instrument hardware [consisting of gel module, computer with mouse and Inkjet-color printer; all necessary components included: power supply, separation chamber, densitometer and computer] - Complete set of software modules [operating and evaluation software] - Palette of accessories: · Gel casting device, gel trays, sample applicators · Concentrator applicator · Additional electrodes for Selective ConcentrationTM (size-specific stacking), In-GelTM Assay, Selective RetrievalTM (zone electroelution) or re-application [multi-dimensional analysis] · Gel format adaptors - Applications Protocols [Procedures for labeling, separation, assays] - reagents, electrolyte buffers, labeling reagents, reference samples - ready-to-use precast gels in Agarose and Polyacrylamide [3 mm preparative gels / 1.5 mm analytical gels]
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