Investigation of voltammetric enzyme-linked immunoassay based on a new system of HAP-H2O2-HRP.
By introducing heterocyclic compound to immunoassay system as an electrochemical substrate for the fist time, a new voltammetric enzyme-linked immunoassay system of 3-hydroxyl-2-aminopyridine (HAP)-H(2)O(2)-horseradish peroxidase (HRP) has been developed. HAP was oxidized with H(2)O(2) catalyzed by HRP, and the resulting electroactive product produced a sensitive voltammetric peak at potential of -0.36 V (vs. SCE) in Britton-Robinson (BR) buffer solution. The process of the enzyme-catalyzed reaction and the electro-reduction of the product have been investigated in detail. The linear range for detection of free HRP was from 4.0x10(-13) to 1.0x10(-9) g/mL with a detection limit of 1.2x10(-13) g/mL. The new system has been successfully applied for the assay of alpha-fetoprotein (alphaFP) in human serum ranging from 0.1 to 200 ng/mL with a detection limit of 0.1 ng/mL, which was 10 times lower than that of traditional spectrophotometric enzyme-linked immunosorbent assay (ELISA) method. HAP-H(2)O(2)-HRP voltammetric enzyme-linked immunoassay showed a promising alternative approach in the detection of alphaFP in clinical diagnosis.
Zhang
Shusheng
S
Key Laboratory of Eco-chemical Engineering, Ministry of Education; College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, China. shushzhang@126.com
Zou
Jin
J
Yu
Fengli
F
D000631:Aminopyridines / Q000737:chemistry*; D002021:Buffers; D002384:Catalysis; D004563:Electrochemistry; D004566:Electrodes; D006735:Horseradish Peroxidase / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques / Q000379:methods*; D010084:Oxidation-Reduction; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D000509:alpha-Fetoproteins / Q000032:analysis
D000631:Aminopyridines; D002021:Buffers; D000509:alpha-Fetoproteins; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; C531569:2-amino-3-hydroxypyridine