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Three Ways Multiplexing Improves Antibody Epitope Mapping

Luminex
8 Dec 2014

Epitope mapping is the process of identifying all of the antibody binding sites on a target protein. Epitope mapping facilitates drug design and patenting, and is therefore one of the key elements in drug development. It is also important for development of antibodies to be used in research or diagnostic assays, which minimizes cross-reactivity. This white paper discusses how epitope mapping can be improved by using Luminex® xMAP® multiplex assays instead of traditional ELISA technology.

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Luminex® 200™Multiplexing Instrument

Luminex

The Luminex® 200™ detects up to 100 protein or nucleic acid analytes from a single aliquot of sample in a single reaction well, enabling you to generate more data while saving sample, time, and reagents. With more than 11,000 units sold globally to date, the Luminex 100/200 is the established standard in multiplexing instrumentation. The xMAP® multiplexing platform is composed of an instrument and associated reagents: The Luminex 200 instrument is based on the principles of flow cytometry, which integrates key xMAP detection components such as lasers, optics, fluidics, and high-speed digital signal processors. xMAP microspheres are a family of fluorescently dyed carboxylated polystyrene microspheres that act as both the surface for the solution phase assay and the spectral identifier that the instrument detects. The open architecture of xMAP Technology enables users to build custom multiplex assays or select from a broad menu commercially available kits. Instrument Details: Multiplex Capacity: Up to 100 non-magnetic; 80 magnetic Microtiter Plate: 96 well Throughput: ~40 min/96-well plate (up to 12,800 tests per hour) Dynamic Range: 3.5 logs Sensitivity: Detects a minimum of 1,000 fluorochromes of phycoerythrin (PE) per xMAP microsphere Optics: Lasers/APDs/PMTs Hardware: Flow Cytometry based Software: xPONENT® Software supports protocol-based data acquisition with robust data regression analysis Dimensions: All components combined (Analyzer, XY Platform and Sheath Delivery System) 26.5" W x 23.5" D x 12.5" H (67.3 cm W x 59.7 cm D x 31.8 cm H) Weight: All components combined (Analyzer, XY Platform and Sheath Delivery System) 113 lbs (49 kg) xMAP Reagent Compatibility: Magnetic and nonmagnetic microspheres

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AntibodiesAntibodies are used in techniques such as confocal and fluorescence microscopy, flow cytometry, ELISA, ELISPOT, immunohistochemistry, western blotting and immunopreciptation. Select specific antigen reactivity, high specific affinity, low non-specific binding, monoclonal or polyclonal, primary or secondary antibodies and associated conjugates such as an enzyme or dye for visualization.MicroplatesMicroplates are multi-well plates used to increase the throughput of biological investigations. The number of wells microplates contain can range from 6 to 3243 wells, with the 96-well format being the most commonly used. Microplates can come tailored for a range of applications including cell culture, PCR, filtration, storage, non-binding surface, protein crystallization, as well as pre-coated, deep well and normal Standard microplates. Additionally, options for microplate colors include clear , black, white or black with clear bottom and white with clear bottom for absorbance microplate reader applications. Find the best microplates for your lab in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Labware and Lab SuppliesLabware and lab supplies include various forms of glassware, plasticware, reagents and personal protective equipment (PPE) and are used across all scientific laboratories, enabling daily tasks to be performed. Explore a range of flasks and beakers, petri dishes, carboys and storage bottles. Labware such as condensers, desiccators and burettes are also available for more specific tasks. Find the best labware and lab supplies in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Microplate Readers / DetectorsMicroplate readers are used to automate the detection and analysis of labeled or label-free components in microplates during assays or live-cell monitoring. Microplate readers are generally distinguished by their mode of detection. Types include absorbance, luminescence, fluorescence intensity, fluorescence polarization, TRF / FRET and multimode microplate readers. Microplate readers deliver a high throughput of samples by reading multiple wells simultaneously, with the 96-well format the most commonly used. As a result, microplate readers are often used in the drug discovery, bioassays, research and pharmaceutical industries for screening applications. Microplate loading can also be automated, with robotic microplate stackers to increase throughput. Find the best microplate readers in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Cell-Based AssaysCell-based assays are used to monitor the presence, quantity and activities of a desired cellular analyte including drug molecules or biomarkers. This can reveal information on cell health (apoptosis, cytotoxicity, viability and proliferation assays), cell metabolism, cell migration and cell signaling mechanisms. Find the best cell-based assay products, kits and equipment with our peer reviewed product directory: compare products, check customer reviews and receiving pricing direct from manufacturers.Automated Liquid HandlingAutomated liquid handling equipment is used to mix, dilute and dispense allotted volumes of liquid into microplates and other vessels automatically. The robotic, liquid handling arms can dispense from single channel to 3456 multichannel as well as operate nanoliter dispensing, enabling high throughput of samples. Find the best automated liquid handlers in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.RegulatoryPharmaceutical regulations impact on all areas of drug development, manufacture and control and supply. Services are provided from the early stages of drug development and clinical trials through to dossier submission, approval and marketing. Regulatory also includes submission of analytical studies from the pharmaceutical product and safety. EpitopeDrug DevelopmentDrug development refers to the process of bringing a new drug to market.Antibody Arrays

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Three Ways Multiplexing Improves Antibody Epitope Mapping