Multiplex Cellular Assays Using Detectable Cell Barcodes

Abstract

We describe herein a cell-based multiplexing technique called detectable cell barcoding (DCB). In DCB, each individual sample is labeled with a different DCB signature that distinguishes each sample by one or both of detected intensity or type of detection characteristic. The samples are then combined and analyzed for a detectable characteristic of interest (e.g., presence of an analyte). By employing multiple distinct DCB labels at varying concentrations, one can perform multiplex analyses on up to hundreds or thousands (or more) of cell samples in a single reaction tube. DCB reduces reagent consumption by factors of 100-fold or more, significantly reduces data acquisition times and allows for stringent control sample analysis.

Description

BACKGROUND OF THE INVENTION[0001]As the scale of cellular analysis experiments expands from a few dozen samples to hundreds or even thousands of individual samples (e.g., in a high-throughput screen), several limitations are encountered, including reagent expense, analysis time, control of assay parameters between different samples, and sample acquisition throughput. Throughput has been increased by rapid auto-samplers, but such systems are not widely available (see, e.g., Kuckuck et al. Cytometry 44, 83-90 (2001); and Edwards et al. Curr. Opin. Chem. Biol. 8, 392-398 (2004), which discuss auto samplers for flow cytometers).[0002]The present invention provides methods and compositions for improving multiple sample (e.g., high throughput) flow cytometric assays.SUMMARY OF THE INVENTION[0003]The present invention is drawn to multiplex cellular assays that employ Detectable Cell Barcodes (DCB). In certain embodiments, different cell samples are labeled with different amounts of a DCB m...

AI Technical Summary

Benefits of technology

[0003]The present invention is drawn to multiplex cellular assays that employ Detectable Cell Barcodes (DCB). In certain embodiments, different cell samples are labeled with different amounts of a DCB marker, e.g., by treatment with different concentrations of a DCB label that binds to a cell (e.g., a cell-reactive form of a fluorophore or a cell-reactive molecular mass marker). This gives each sample a unique signature upon analysis (e.g., flow cytometric detection and / or mass spectrometer analysis). In certain embodiments, cell samples are coded with more than one DCB marker (e.g., DCB markers having distinct detection characteristics). In these embodiments, the number of different DCB signatures available increases geometrically because of multiplexing of DCB intensity with DCB detection characteristic (for example, using different channels on the flow cytometer). DCB allows the multiplex analysis of hundreds to thousands of samples (or more) in a single reaction tube, which significantly reduces regent consumption, improves the throughput of experiments, and eliminates potential sample to sample variability.

Problems solved by technology

As the scale of cellular analysis experiments expands from a few dozen samples to hundreds or even thousands of individual samples (e.g., in a high-throughput screen), several limitations are encountered, including reagent expense, analysis time, control of assay parameters between different samples, and sample acquisition throughput.

Throughput has been increased by rapid auto-samplers, but such systems are not widely available (see, e.g., Kuckuck et al.

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