Kinetics study on recombinant alkaline phosphatase and correlation with the fluorescent signal generated.

Kinetics study on recombinant alkaline phosphatase and correlation with the fluorescent signal generated. J Immunoassay Immunochem. 2017 Dec 15;: Authors: Susini V, Rossi VL, Sanesi A, Drazek L Abstract Alkaline phosphatase (AP) (EC 3.1.3.1) is one of the most used enzymes in immunoassays. In VIDAS® assays (bioMérieux, Marcy l'Etoile, France) AP catalyses the hydrolysis of 4-Methylumbelliferyl phosphate (4-MUP) in 4-Methylumbelliferone (4-MU) producing a fluorescent signal. This work introduces an original method of characterization of the kinetics parameter Km, Vmax and Kcat of AP embedded in VIDAS® assays. Assessment of such constants allows us to predict the fluorescent signal generated for given amounts of enzyme and its associated substrate; in the particular case of VIDAS®, it has been estimated that 0.06 nmol/L of AP produce 3144 Relative Fluorescent Values (RFV). PMID: 29244613 [PubMed - as supplied by publisher]
Source: Journal of Immunoassay and Immunochemistry - Category: Biochemistry Tags: J Immunoassay Immunochem Source Type: research