Clinical Chemistry Siemens Point of Care - Urinalysis
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Clinical Chemistry 43: 2128-2136, 1997;
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(Clinical Chemistry. 1997;43:2128-2136.)
© 1997 American Association for Clinical Chemistry, Inc.

Articles

Fiber-optic immunosensor for measurement of myoglobin

C. Michael Hanbury1,3, W. Greg Miller1,a and Robert B. Harris2

Departments of
1 Pathology and
2 Biochemistry and Molecular Biophysics, Medical College of Virginia, Virginia Commonwealth University, Richmond, VA 23298-0597.
3 Current address: Roche Diagnostics, 1080 US Highway 202, Somerville, NJ 08876.
a Author for correspondence: Medical College of Virginia, Virginia Commonwealth University, P.O. Box 980286, Richmond, VA 23298-0286. Fax 804-828-0353; e-mail millerg{at}gems.vcu.edu

A self-contained fiber-optic immunosensor was developed to measure the 16 500-Da protein myoglobin. The sensing element was constructed by entrapment of Cascade Blue-labeled antibody within polyacrylamide gel at the distal face of an optical fiber 300 µm in core diameter. The polyacrylamide gel composition was optimized to allow diffusion of myoglobin but to exclude hemoglobin and higher-molecular-mass proteins from the sensing area. The analytical signal was derived from fluorescence energy transfer between Cascade Blue and the heme group of myoglobin. Fluorescence quenching occurred when myoglobin bound to labeled antibody. The total amount of fluorescence quench was dependent on the antibody labeling conditions and the amount of antibody incorporated in the sensor gel matrix. Myoglobin concentrations >5 nmol/L (83 µg/L) were measurable with response times of 15 to 130 min limited by diffusion into the sensing element. This report demonstrates the technical feasibility for a self-contained immunosensor to measure a protein analyte.






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Copyright © 1997 by the American Association for Clinical Chemistry.