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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 25, Number 1
BKCSDE 25(1)
January 20, 2004 

Homogeneous Enzyme-Linked Binding Assay Mediated by the Interaction of Avidin with Biotin: Mistletoe Lectin I Assay
Insook Rhee Paeng*, Eun Ah Lee, Hyun Sook Kim
ELBA, Avidin/Biotin, ML I, Homogeneous assay
We have examined the feasibility of using the specific interaction between mistletoe lectin I (ML I) and β-Dgalactose instead of the anti-ML I antibody in developing a homogeneous type competitive binding assay for ML I. We also have examined the feasibility of adapting the biotin/avidin mediated homogeneous assay for this system. Alkaline phosphatase (AKP) was employed as a single substrate enzyme label. The dose-response curve shows a detection range of 1-25 μg/mL and a linear response with a correlation coefficient of 0.99. To demonstrate the analytical utility of this method, 10 μg/mL of ML I was spiked into distilled water. The results show that the mean recovery was 10.03 μg/mL with an SD of 0.18. The difference between the spiked value and the mean recovery was 0.03 μg/mL, with a relative error of 0.3 and 1.6 % of RSD.
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