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Published February 21, 2007 | Accepted Version + Supplemental Material
Journal Article Open

DNA-Encoded Antibody Libraries: A Unified Platform for Multiplexed Cell Sorting and Detection of Genes and Proteins


Whether for pathological examination or for fundamental biology studies, different classes of biomaterials and biomolecules are each measured from a different region of a typically heterogeneous tissue sample, thus introducing unavoidable sources of noise that are hard to quantitate. We describe the method of DNA-encoded antibody libraries (DEAL) for spatially multiplexed detection of ssDNAs and proteins as well as for cell sorting, all on the same diagnostic platform. DEAL is based upon the coupling of ssDNA oligomers onto antibodies which are then combined with the biological sample of interest. Spotted DNA arrays, which are found to inhibit biofouling, are utilized to spatially stratify the biomolecules or cells of interest. We demonstrate the DEAL technique for (1) the rapid detection of multiple proteins within a single microfluidic channel, and, with the additional step of electroless amplification of gold-nanoparticle labeled secondary antibodies, we establish a detection limit of 10 fM for the protein IL-2, 150 times more sensitive than the analogue ELISA; (2) the multiplexed, on-chip sorting of both immortalized cell lines and primary immune cells with an efficiency that exceeds surface-confined panning approaches; and (3) the co-detection of ssDNAs, proteins, and cell populations on the same platform.

Additional Information

© 2007 American Chemical Society. Received August 15, 2006. Publication Date (Web): January 30, 2007. The authors thank Leroy Hood, Paul Mischel, and Toni Ribas for helpful discussions. We also thank Bruz Marzolf at the Institute for Systems Biology (Seattle, WA) for printing DNA microarrays and Mireille Riedinger at UCLA for outstanding technical assistance with primary cell purifications. The VL-3 T cells were a kind gift from Dr. Stephen Smale at UCLA. We thank Harry Choi and Suvir Venkataraman in the Pierce group at Caltech for their help in generating the orthogonal DNA sequences. This work was funded by the National Cancer Institute Grant No. 5U54 CA119347 (J.R.H., P.I.) and by the DOE-funded Institute for Molecular Medicine Laboratory. Owen Witte is an Investigator of the Howard Hughes Medical Institute.

Attached Files

Accepted Version - nihms-61976.pdf

Supplemental Material - ja065930isi20061128_112719.pdf


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