The BD Cytometric Bead Array System (CBA)
Table of Contents
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3
A Multiplex Bead System You Can Count On . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3
BD Cytometric Bead Array (CBA) Assay Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4
Quality is Built in to Every BD CBA Product . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5
Standardization of BD CBA Standards to International (NIBSC) Standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9
Non-human Primate Cross-reactivity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11
BD FACSArray Bioanalyzer is Ideal for BD CBA Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12
BD CBA Assay Troubleshooting Tips . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .13
Product Listing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14
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Microsoft and Windows are registered trademarks of Microsoft Corporation.For Research Use Only. Not for use in diagnostic or therapeutic procedures.All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.Purchase does not include or carry any right to resell or transfer this product either as a stand-alone product or as a component of another product. Any use of this product other than the permitted use without the express written authorization of Becton Dickinson and Company is strictly prohibited.BD flow cytometers are class I (1) laser productsBD, BD Logo and all other trademarks are the property of Becton, Dickinson and Company. 2004 BD
A Multiplex Bead System You Can Count On
BD Biosciences has added a new twist to counting with beads. Werecognize the value of precious samples and data reproducibility,so weve developed a technology to ensure both. The innovativeBD Cytometric Bead Array (CBA) technology allows forquantitative detection of multiple analytes in a single serum,plasma, tissue culture supernatant, or cell lysate sample. TheBD CBA System of assay kits, flow cytometers, and easy-to-usesoftware provides reproducible data and reliable performance thatyou can count on time and time again.
Introduction
Unless otherwise specified, all products are for Research Use Only.Not for use in diagnostic or therapeutic procedures. Not for resale.
Get multiple results from a singlesmall-volume sample
Run one standard mixture to generatestandard curves for all your analytes
Avoid artifacts associated withenzyme-dependent signal generation
Achieve quantitative results with lesstime and labor
Combine versatile flow cytometerswith ready-to-use kits and analysissoftware
Automate sample acquisition andincrease throughput with the plate-based BD FACSArray bioanalyzer orother BD flow cytometers equippedwith the high throughput sampler(HTS) option.
With the BD Cytometric Bead Array(CBA) System You Can:
Flow cytometry is an analytical tool that allows for thediscrimination of different particles on the basis of size andcolor. The BD CBA employs a series of particles with discretefluorescence intensities to simultaneously detect multiple solubleanalytes from a single serum, plasma, or tissue culturesupernatant sample. The BD CBA, combined with flow cytometry,creates a powerful multiple analyte (multiplex) assay system. TheBD CBA system uses the sensitivity of amplified fluorescencedetection by flow cytometry to measure soluble analytes with aparticle-based immunoassay. The combined advantages of thebroad dynamic range of fluorescence detection via flowcytometry, and the efficient capturing of analytes via suspendedparticles coated with distinct capture antibodies enable theBD CBA to use fewer sample dilutions to determine analyteconcentration in substantially less time (compared toconventional ELISA).
The specific capture beads are mixed with the phycoerythrin (PE)conjugated detection antibodies and then incubated withrecombinant protein standards or test samples to form sandwichcomplexes. Following acquisition of sample data using the flowcytometer, the sample results are generated in graphical andtabular format using the BD CBA Analysis Software.
BD CBA Assay Overview
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Not for use in diagnostic or therapeutic procedures. Not for resale.
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W A S H
I N C U B A T E
A N A L Y Z E
A C Q U I R E
BD CBAFigure 1. Typical BD CBA assay protocol. A singleanalyte is shown for representational purposes.
The powerful BD Cytometric BeadArray (CBA) assays enable multiplexanalysis of complex biological sampleson a flow cytometer. Each assay has beenstringently developed for ease-of-use,broad instrument compatibility and rapiddata analysis, sensitivity, reproducibility,and quality.
As with any complex assay, the ability ofthe novice user to become comfortablewith the assay is critical. In the BD CBAsystem, ease-of-use has been engineeredinto each and every kit. The standards foreach multiplex kit are provided togetherin a lyophilized format that yields a ready-to-use stock standard solution uponreconstitution. Each kit includes a pre-diluted detection reagent containing eachdetector antibody formulated at theiroptimal concentration. Even the assayprotocol has been designed to limithandling steps (Figure 1) and hands-ontime by allowing the researcher to add allkit reagents, samples, and/or standards totheir assay well or tube at the same time.The assay is incubated and thencompleted with a short wash stepfollowed by sample analysis on a flowcytometer.
Every BD CBA assay is compatible withany flow cytometer that is equipped witha 488 nm laser and capable of detectingand distinguishing fluorescence emissionsat 576 and 670 nm. In addition, analysissoftware capable of saving data in an FCS
2.0 file format is required. This includesall of the BD flow cytometry systems,however, it should be noted that stream-in-air instruments will yield lower assaysensitivities than other instruments. Thedata collected on a flow cytometer, oncesaved as an FCS 2.0 file, can be rapidlyanalyzed using the BD CBA Software.The BD CBA Software enables linearregression analysis of data files andextrapolation of sample values bycomparison against a known standardcurve. The software also has broadcompatibility as it is offered in both PCand Mac-compatible formats, requiringonly Microsoft Excel to run. With theBD CBA Software, sample results areobtained within minutes of completing anexperiment.
Considerable effort has been made toensure that each BD CBA kit has thehighest possible sensitivity and bestreproducibility. Each antibody pair used inthe kits is evaluated for dynamic range,sensitivity, and parallel titration curves tonative biological samples (Figures 2and 3). In addition, the scientists atBD Biosciences have formulated the assaydiluent and wash buffers in each kit toreduce detrimental effects of serum andplasma proteins on assay performance.While this does not always yieldcomparable recovery results as singleassays (eg, ELISA), it does provide qualitymultiplex performance.
Possibly the most important aspect of theBD CBA kits is the quality performancethat is built into each one. Usingtechniques and methods developed atBD Biosciences, a worldwide leader in theproduction and conjugation of antibodies,the reagents in a BD CBA kit arestringently tested during the developmentprocess. Each capture antibody used for acapture bead has been tested both beforeand after it is coupled to the bead.Further, it is tested again when it iscombined with the other kit reagents tocomplete the manufacture of a batch ofkits. The same is true of the detectionantibodies, which are additionally testedbefore and after conjugated withR-phycoerythrin (R-PE) and again whenthey are formulated into a detectionreagent. Finally, they are tested with theother reagents in a given kit batch afterthey are bottled. All of this effort is madeto ensure that the kit that is delivered tothe researcher meets their expectationsand provides equal performance everytime. Further, the quality of the BD CBAkits does not end with the release of theproduct. We are continually looking formethods to enhance kit performancethrough improved antibody pairs, newstandards formulations and optimizedbuffers for serum samples.
Quality is Built into Every BD CBA Product
5Unless otherwise specified, all products are for Research Use Only.Not for use in diagnostic or therapeutic procedures. Not for resale.
IL-12p70
TNFIL-10 IL-6
IL-1
IL-8
Figure 2a. Representative data generated using the BD CBA Human Inflammation Kit.Relative bead fluorescence intensities.
Quality is Built into Every BD CBA Product
6 www.bdbiosciences.comUnless otherwise specified, all products are for Research Use Only.
Not for use in diagnostic or therapeutic procedures. Not for resale.
Figures 2c. Representative data generated using the BD CBA Human Inflammation Kit.Typi
