Abstract
As an important DNA 3′-phosphatase, alkaline phosphatase can repair damaged DNA caused by replication and recombination. It is essential to measure the level of alkaline phosphatase to indicate some potential diseases, such as cancer, related to alkaline phosphatase. Here, we designed a simple and fast method to detect alkaline phosphatase quantitively. When alkaline phosphatase is present, the resulting poly T-DNA with a 3′-hydroxyl end was cleaved by exonuclease I, prohibiting the formation of fluorescent copper nanoparticles. However, the fluorescent copper nanoparticles can be monitored with the absence of alkaline phosphatase. Hence, we can detect alkaline phosphatase with this turn-off strategy. The proposed method is able to quantify the concentration of alkaline phosphatase with the LOD of 0.0098 U/L. Furthermore, we utilized this method to measure the effects of inhibitor Na3VO4 on alkaline phosphatase. In addition, it was successfully applied to quantify the level of alkaline phosphatase in human serum. The proposed strategy is sensitive, selective, cost effective, and timesaving, having a great potential to detect alkaline phosphatase quantitatively in clinical diagnosis.
Highlights
Alkaline phosphatase (ALP), a ubiquitous enzyme found in human tissues such as the liver, intestine, bone, kidney, and placenta, is a homodimeric enzyme with necessary cofactors, including one magnesium atom and two zinc atoms [1,2]
Considering that poly T DNA, an ideal template of fluorescent CuNPs, can be modified by the removal of a phosphate group from the 3 -end catalyzed by ALP, we have proposed a method for the measurement of ALP concentration with LOD of 0.0098 U/L requiring only 50 min based on terminal protection and fluorescent CuNPs
AHLoPw, esvuecrh, tahserdeiaabreetsetsil,lbsroemasetcchaanlcleenr,gaens dtoporvoesrtcaotimc ecainncperraccltiincaiclaalplyp[li9c–a1t1i]o.nHs.oFwoerveexra,mthpelree, tahries mstieltlhsoodmreeqcuhiarlelsenagdeisffteoreonvterrecaocmtioeninbupfrfearctwichailcahpips laiccahtiaollnesn.gFeoirn epxaramctpiclea,ltahpispmliceatthioondsr. equires a different reaction buffer which is a challenge in practical applicIanticoonnsc. lusion, based on the poly T-DNA-templated formation of fluorescent CuNPs, we hInavceonpcrloupsoiosne,dbaasfeadciolenbtuhtespeonlsyitTiv-De,NseAle-tcetmivpe,lalotewd-fcoorsmt,aatniodntoimf felu-soarveisncgenAtLCPuNasPsas,y. we have proposed a facile but sensitive, selective, low-cost, and time-saving ALP assay
Summary
Alkaline phosphatase (ALP), a ubiquitous enzyme found in human tissues such as the liver, intestine, bone, kidney, and placenta, is a homodimeric enzyme with necessary cofactors, including one magnesium atom and two zinc atoms [1,2]. Owing to the indispensable role in many physiological processes such as cell cycle, growth, apoptosis, and signal transduction, ALP is closely connected to multiple human diseases, especially bone and hepatic diseases [5,6,7]. Any abnormal level of ALP in the serum may be an essential indicator of some diseases related to ALP function, such as diabetes, breast cancer, prostatic cancer, bone diseases, such as osteosarcoma, and hepatic diseases, e.g., Wilson’s disease. ALP levels in the serum may be an effective biomarker in medical diagnosis [9,10,11]. Prakash et al have recently found that the level of ALP in saliva (readily accessible, safe, and noninvasive body fluid) may be able to serve as an early biomarker for diabetes mellitus and some potentially malignant tumors [12]. It is of great importance to developing a facile and sensitive method to detect ALP
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