AquaLite, a bioluminescent label for immunoassay and nucleic acid detection: quantitative analyses at the attomol level

David F. Smith; Nancy L. Stults

doi:10.1117/12.237603

1 April 1996 AquaLite, a bioluminescent label for immunoassay and nucleic acid detection: quantitative analyses at the attomol level

David F. Smith, Nancy L. Stults

Author Affiliations +

Proceedings Volume 2680, Ultrasensitive Biochemical Diagnostics; (1996) https://doi.org/10.1117/12.237603
Event: Photonics West '96, 1996, San Jose, CA, United States

Abstract

AquaLite^R is a direct, bioluminescent label capable of detecting attomol levels of analyte in clinical immunoassays and assays for the quantitative measurement of nucleic acids. Bioluminescent immunoassays (BIAs) require no radioisotopes and avoid complex fluorescent measurements and many of the variables of indirect enzyme immunoassays (EIAs). AquaLite, a recombinant form of the photoprotein aequorin from a bioluminescent jellyfish, is coupled directly to antibodies to prepare bioluminescent conjugates for assay development. When the AquaLite-antibody complex is exposed to a solution containing calcium ions, a flash of blue light ((lambda) _max equals 469 nm) is generated. The light signal is measured in commercially available luminometers that simultaneously inject a calcium solution and detect subattomol photoprotein levies in either test tubes or microtiter plates. Immunometric or 'sandwich' type assays are available for the quantitative measurement of human endocrine hormones and nucleic acids. The AquaLite TSH assay can detect 1 attomol of thyroid stimulating hormone (TSH) in 0.2 mL of human serum and is a useful clinical tool for diagnosing hyperthyroid patients. AquaLite-based nucleic acid detection permits quantifying attomol levels of specific nucleic acid markers and represents possible solution to the difficult problem of quantifying the targets of nucleic acid amplification methods.

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David F. Smith and Nancy L. Stults "AquaLite, a bioluminescent label for immunoassay and nucleic acid detection: quantitative analyses at the attomol level", Proc. SPIE 2680, Ultrasensitive Biochemical Diagnostics, (1 April 1996); https://doi.org/10.1117/12.237603

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