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Clinical Chemistry 47: 1894-1900, 2001;
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(Clinical Chemistry. 2001;47:1894-1900.)
© 2001 American Association for Clinical Chemistry, Inc.

Oak Ridge Conference

Silicon-based Biosensors for Rapid Detection of Protein or Nucleic Acid Targets

Robert Jenison1, Helen La1, Ayla Haeberli1, Rachel Ostroff1 and Barry Polisky1a

1 ThermoBioStar, Inc., 6655 Lookout Rd., Boulder, CO 80301.

aAuthor for correspondence. Fax 303-581-6405; e-mail b_polisky{at}thermobiostar.com.


Abstract

Background: We developed a silicon-based biosensor that generates visual, qualitative results or quantitative results for the detection of protein or nucleic acid targets in a multiplex format.

Methods: Capture probes were immobilized either passively or covalently on the optically coated surface of the biosensor. Intermolecular interactions of the immobilized capture probe with specific target molecules were transduced into a molecular thin film. Thin films were generated by enzyme-catalyzed deposition in the vicinity of the surface-bound target. The increased thickness on the surface changed the apparent color of the biosensor by altering the interference pattern of reflected light.

Results: Cytokine detection was achieved in a 40-min multiplex assay. Detection limits were 4 ng/L for interleukin (IL)-6, 31 ng/L for IL1-ß, and 437 ng/L for interferon-{gamma}. In multianalyte experiments, cytokines were specifically detected with signal-to-noise ratios ranging from 15 to 80. With a modified optical surface, specificity was also demonstrated in a nucleic acid array with unambiguous discrimination of single-base changes in a 15-min assay. For homozygous wild-type and homozygous mutant samples, signal-to-noise ratios of ~100 were observed. Heterozygous samples yielded approximately equivalent signals for wild-type and mutant capture probes.

Conclusions: The thin-film biosensor allows rapid, sensitive, and specific detection of protein or nucleic acid targets in an array format with results read visually or quantified with a charge-coupled device camera. This biosensor is suited for multianalyte detection in clinical diagnostic assays.




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X.-b. Zhong, R. Reynolds, J. R. Kidd, K. K. Kidd, R. Jenison, R. A. Marlar, and D. C. Ward
Single-nucleotide polymorphism genotyping on optical thin-film biosensor chips
PNAS, September 30, 2003; 100(20): 11559 - 11564.
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