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Diffractive Micro Bar Codes for Encoding of Biomolecules in Multiplexed Assays

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Graham R. Broder,^† Rohan T. Ranasinghe,^† Joseph K. She,^† Shahanara Banu,^†^‡ Sam W. Birtwell,^§^‖ Gabriel Cavalli,^† Gerasim S. Galitonov,^§ David Holmes,^‡ Hugo F. P. Martins,^† Kevin F. MacDonald,^§ Cameron Neylon,^† Nikolay Zheludev,^§ Peter L. Roach,*^† and Hywel Morgan*^‡

School of Chemistry, School of Electronics and Computer Science, Optoelectronics Research Centre, and School of Physics and Astronomy, University of Southampton, Highfield, Southampton, SO17 1BJ, UK, and STFC, Rutherford Appleton Laboratory, Didcot, OX11 0QX, UK

Anal. Chem., 2008, 80 (6), pp 1902–1909

DOI: 10.1021/ac7018574

Publication Date (Web): February 14, 2008

Section:

Biochemical Methods

Abstract

Microparticles incorporating micrometer-sized diffractive bar codes have been modified with oligonucleotides and immunoglobulin Gs to enable DNA hybridization and immunoassays. The bar codes are manufactured using photolithography of a chemically functional commercial epoxy photoresist (SU-8). When attached by suitable linkers, immobilized probe molecules exhibit high affinity for analytes and fast reaction kinetics, allowing detection of single nucleotide differences in DNA sequences and multiplexed immunoassays in <45 min. Analysis of raw data from assays carried out on the diffractive microparticles indicates that the reproducibility and sensitivity approach those of commercial encoding platforms. Micrometer-sized particles, imprinted with several superimposed diffraction gratings, can encode many million unique codes. The high encoding capacity of this technology along with the applicability of the manufactured bar codes to multiplexed assays will allow accurate measurement of a wide variety of molecular interactions, leading to new opportunities in diverse areas of biotechnology such as genomics, proteomics, high-throughput screening, and medical diagnostics.

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