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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 17, Number 11
BKCSDE 17(11)
November 20, 1996 

Potentiometric Homogeneous Enzyme-Linked Binding Assays for Riboflavin and Riboflavin Binding Protein
Jin Mog Kim, Hye Jin Kim, Mi Jeoung Kim, Dong Joo Lee, Sang Hyun Han, Geun Sig Cha*
Adenosine deaminase (ADA) has been utilized as the label in devising a potentiometric homogeneous assay for riboflavin and riboflavin binding protein (RBP). The proposed homogeneous assay method employs an ADA-biotin conjugate as the signal generator and an avidin-riboflavin conjugate as the signal modulator in the solution phase. The catalytic activity of the ADA-biotin conjugate is inhibited in the presence of an excess amount of the avidin-riboflavin conjugate, and the observed inhibition is reversed in an amount proportional to the concentration of RBP added. When the analyte riboflavin is added to this mixture of ADA-biotin, avidin-riboflavin and RBP, the activity of the enzyme conjugate is re-inhibited in an amount proportional to the concentration of riboflavin. Since the enzyme label used in this system is ADA, an ammonia-producing enzyme, a potentiometric rather than photometric detection scheme is used to monitor the enzymatic activity in the assay.
1018 - 1022
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