title: Sensitive label-free electrochemical analysis of human IgE using an aptasensor with cDNA amplification.
abstract: In this study, we developed an ultrasensitive label-free aptamer-based electrochemical biosensor, featuring a highly specific anti-human immunoglobulin E (IgE) aptamer as a capture probe, for human IgE detection. Construction of the aptasensor began with the electrodeposition of gold nanoparticles (AuNPs) onto a graphite-based screen-printed electrode (SPE). After immobilizing the thiol-capped anti-human IgE aptamer onto the AuNPs through self-assembly, we treated the electrode with mercaptohexanol (MCH) to ensure that the remaining unoccupied surfaces of the AuNPs would not undergo nonspecific binding. We employed a designed complementary DNA featuring a guanine-rich section in its sequence (cDNA G1) as a detection probe to bind with the unbound anti-human IgE aptamer. We measured the redox current of methylene blue (MB) to determine the concentration of human IgE in the sample. When the aptamer captured human IgE, the binding of cDNA G1 to the aptamer was inhibited. Using cDNA G1 in the assay greatly amplified the redox signal of MB bound to the detection probe. Accordingly, this approach allowed the linear range (coefficient of determination: 0.996) for the analysis of human IgE to extend from 1 to 100,000pM; the limit of detection was 0.16pM. The fabricated aptasensor exhibited good selectivity toward human IgE even when human IgG, thrombin, and human serum albumin were present at 100-fold concentrations. This method should be readily applicable to the detection of other analytes, merely by replacing the anti-human IgE aptamer/cDNA G1 pair with a suitable anti-target molecule aptamer and cDNA.
authors.affiliation: Department of Applied Chemistry, National Chiao Tung University, Hsinchu, Taiwan, ROC.
authors.forename: Cheng-Yu
authors.initials: CY
authors.lastname: Lee
authors.forename: Kuan-Ying
authors.initials: KY
authors.lastname: Wu
authors.forename: Hsiu-Li
authors.initials: HL
authors.lastname: Su
authors.forename: Huan-Yi
authors.initials: HY
authors.lastname: Hung
authors.forename: You-Zung
authors.initials: YZ
authors.lastname: Hsieh
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods; D018076:DNA, Complementary / Q000737:chemistry; D055664:Electrochemical Techniques / Q000379:methods*; D004566:Electrodes; D006046:Gold / Q000737:chemistry; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis*; D057230:Limit of Detection; D053768:Metal Nanoparticles / Q000737:chemistry; D013917:Thrombin / Q000032:analysis
chemical_list: D052157:Aptamers, Nucleotide; D018076:DNA, Complementary; D007073:Immunoglobulin E; D006046:Gold; D013917:Thrombin