Key Laboratory of Chemical Biology & Traditional Chinese Medicine Research, Ministry of Education, Hunan Normal University, Changsha 410081, China.
Analyst. 2014 Jan 7;139(1):285-9. doi: 10.1039/c3an01736b. Epub 2013 Nov 19.
Based on the fluorescence quenching of novel denatured protein-protected gold nanoclusters, a label-free detection method of acetylcholinesterase (AChE) activity has been developed. Using denatured bovine serum albumin (dBSA), in which 35 cysteine residues can interact polyvalently with Au nanoclusters (AuNCs) as a stabilizing agent, water-soluble and stable fluorescent gold nanoclusters were synthesized. The fluorescence of the AuNCs was quenched by thiocholine that was produced from the AChE hydrolysis of S-acetylthiocholine iodide (ACTI) to detect the AChE activity. The linear range of the method was 0.005-0.15 U mL(-1). The limit of detection (LOD) was 0.02 mU mL(-1). Other enzymes and metal ions, i.e., GPT, γ-GT, GOx, K(+), Ca(2+) and Na(+), showed minimal interference. Using the fluorescence probe, satisfactory results for the detection of the AChE activity in human serum were obtained.
基于新型变性蛋白保护的金纳米簇的荧光猝灭，建立了一种无需标记即可检测乙酰胆碱酯酶（AChE）活性的方法。使用变性牛血清白蛋白（dBSA）作为稳定剂，其中 35 个半胱氨酸残基可以与 Au 纳米簇（AuNCs）多价相互作用，合成了水溶性和稳定的荧光金纳米簇。AuNCs 的荧光被硫代胆碱猝灭，硫代胆碱是由 AChE 水解 S-乙酰硫代胆碱碘化物（ACTI）产生的，从而可以检测 AChE 的活性。该方法的线性范围为 0.005-0.15 U mL(-1)。检测限（LOD）为 0.02 mU mL(-1)。其他酶和金属离子，如 GPT、γ-GT、GOx、K(+)、Ca(2+)和 Na(+)，几乎没有干扰。使用荧光探针，对人血清中 AChE 活性的检测得到了令人满意的结果。
