12th International Symposium on Bioluminescence & Chemiluminescence

Symposium abstracts:

Single molecule detection of alkaline phosphatase enzyme

Giri, Brij P.¹, Toben, Howard R.¹, Dagli, Dinesh J.¹, Lamkin, Mark S.², Shinefeld, Lisa A.²

(1) Michigan Diagnostics L.L.C., 1197 Rochester Road, Suite L, Troy, MI 48083, USA
(2) Intergen Company, 25 Birch Street, Milford, MA 01757, USA

A detection system for finding the presence of alkaline phosphatase enzyme at attogram level of at a single molecule level in aqueous buffer. A detection system comprising of (1) a stable 1,2-dioxetane derived from the photo-oxidation of novel alkenes which were synthesized by a coupling reaction of a substituted aromatic ester and a spiro-fused ketone with carbon-carbon double bond in the presence of titanium zero, (2) a polymeric enhancer which increases the output of emitted light from the 1,2-dioxetane when triggered by an enzyme, and (3) an enzyme diluent which stabilizes the enzyme during the reaction has been developed. Utilizing the present system for the detection of alkaline phosphatase conjugated probes will be useful for early discovery of infectious bacterial or viral disease or cancer.

This is a preprint of an article accepted for publication in Luminescence: Copyright 2001 John
Wiley & Sons, Ltd (Wiley website)