Sensitive detection of alkaline phosphatase based on terminal deoxynucleotidyl transferase and endonuclease IV-assisted exponential signal amplification.

ABSTRACT: Alkaline phosphatase (ALP) is widely expressed in human tissues. ALP plays an important role in the dephosphorylation of proteins and nucleic acids. Therefore, quantitative analysis of ALP plays a vital role in disease diagnosis and the development of biological detection methods. Terminal deoxynucleotidyl transferase (TdT) catalyzes continuous polymerization of deoxynucleotide triphosphates at the 3'-OH end of single-stranded DNA in the absence of a template. In this study, we developed a highly sensitive and selective method based on TdT and endonuclease IV (Endo IV) to quantify ALP activity. After ALP hydrolyzes the 3'-PO₄ end of the substrate and generates 3'-OH, TdT can effectively elongate the 3'-OH end with deoxynucleotide adenine triphosphate (dATP) and produce a poly A

SUBMITTER: Ye W 

PROVIDER: S-EPMC9463482 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature