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Direct Detection of Isolevuglandins in Tissues Using a D11 scFv-Alkaline Phosphatase Fusion Protein and Immunofluorescence
Published on: July 5, 2021
Mimicking biological process to detect alkaline phosphatase activity using the vitamin B6 cofactor conjugated bovine
Yachana Upadhyay1, Shilpa Bothra1, Rajender Kumar1
1Department of Applied Chemistry, SV National Institute of Technology (SVNIT), Surat, 395007, India.
Abstract:
This manuscript presents a novel bioanalytical approach for the selective ratiometric fluorescent sensing of enzymatic activity of the alkaline phosphatase (ALP) in the biological samples. The probe was designed by conjugating the pyridoxal 5'-phosphate (PLP) over the surface of bovine serum albumin (BSA) stabilized CdS quantum dots (QDs) through the interaction of free amine present in BSA with the aldehyde group of PLP. The conjugation of PLP quenched the emission of QDs. Upon addition of the ALP, the emission of QDs was restored due to the dephosphorylation and the conversion of the functionalized PLP in to pyridoxal. With this probe, the ALP activity can be detected down to 0.05 U/L and also successfully applied for the detection of ALP activity in biological samples such as human serum and plasma.
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