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Saturday, August 19, 2017

All about FPIA Fluorescence Polarization Immunoassay



• Fluorescein-labeled drug competes with unlabeled drug for antibody
•Sample excited with plane polarized light (490 nm)
•Fluorescein emits plane polarized light (520 nm)
•Small, free drug-fluorescein, rotates faster leading to less emission
•Larger, antibody-drug-fluorescein, rotates slower and emits plane polarized light (520 nm)


Principle: Drug in sample competes for antibody with fluorescein bound drug

•More drug in the sample; less fluorescein labeled drug bound to antibody; lower
emission of plane polarized light.

•Higher drug concentration results in lower light emission values.

FPIA
•Available for a variety of drugs

Advantages

Rapid turnaround times, sensitivity, ease of operation

Disadvantages

Background interference in serum sample (requires blank measurement).



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