A rate turbidimetric immunoassay for theophylline using biotin-avidin system

We describe a biotin-avidin based rate turbidimetric homogeneous immunoassay for the determination of serum theophylline with a measuring range of 0-40 micrograms/ml. The assay features an avidin-biotin-labeled theophylline conjugate and a monoclonal antibody. The rate of change of turbidity caused by the antibody-conjugate complexing was monitored on the Beckman Synchron CX 4 System at 340 nm and 37 degrees C. Theophylline in a sample inhibited the complexation, and the extent of inhibition allowed the quantitation of theophylline in the sample. The within-run coefficient of variation (CV) was < 3.7% and the between-run CV was 5.2-9.2% depending on the theophylline concentration. Linear regression analysis showed a good correlation with an established fluorescence polarization immunoassay (r = 0.9866, n = 94).