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

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Contribution from the NanoSystems Biology Cancer Center: Division of Chemistry and Chemical Engineering, MC 127-72, California Institute of Technology, Pasadena, California 91125, and Department of Molecular and Medical Pharmacology, Departments of Microbiology, Immunology, and Molecular Genetics, and Howard Hughes Medical Institute, University of California, Los Angeles, California 90095
Cite this: J. Am. Chem. Soc. 2007, 129, 7, 1959–1967
Publication Date (Web):January 30, 2007
https://doi.org/10.1021/ja065930i
Copyright © 2007 American Chemical Society

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    Abstract

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    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.

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     California Institute of Technology.

     Current address:  Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801.

     Department of Molecular and Medical Pharmacology, University of California.

     Howard Hughes Medical Institute, University of California.

    §

     Departments of Microbiology, Immunology, and Molecular Genetics, University of California.

    *

    In papers with more than one author, the asterisk indicates the name of the author to whom inquiries about the paper should be addressed.

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    Complete refs 4, 7, 8, and 11; DNA sequences; protein arrays Figures S1−S4. This material is available free of charge via the Internet at http://pubs.acs.org.

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