1.Magnetic Affinity Immunoassay Based Enzyme-Labeled Phage Displayed Antibody
Xihui MU ; Zhaoyang TONG ; Qibin HUANG ; Bing LIU ; Zhiwei LIU ; Lanqun HAO ; Jinping ZHANG
Chinese Journal of Analytical Chemistry 2014;(6):785-790
A new magnetic affinity immunoassay (MAIA) strategy based on enzyme-labeled phage displayed antibody was developed. The assay consisted of a sandwich format in which immobilized polyclonal antibody (pcAb) on magnetic microparticle was used for capture probe, and enzyme-labeled phage displayed antibody for specific detection probe to increase enzyme amount and enhance detection signal. By the proposed method,β-bungarotoxin (β-BGT) was successfully detected. A linear relationship between absorbance value and the concentration of β-BGT in the range of 0. 016-62. 5 μg / L was obtained. The linear regression equation was Y=0. 641X+1. 355 (R =0. 9925, n = 13, p<0. 0001) with a detection limit of 0. 016 μg / L. In comparison with the traditional ELISA, this method gave a 10-fold better sensitivity in β-BGT detection. This strategy also gave a 4-fold better sensitivity comparing with the MAIA based on enzyme labeled monoclonal antibody (mcAb). Due to low detection limit, acceptable reproducibility and high specificity, this method holds great promise in toxin trace detection.
2.Aptamer-Based Microcantilever Sensor for O-ethyl S-[2-(diisopropylamino) ethyl] methylphosphonothiolate, Sarin Detection and Kinetic Analysis
Zhiwei LIU ; Zhaoyang TONG ; Lanqun HAO ; Bing LIU ; Xihui MU ; Jinping ZHANG ; Chuan GAO
Chinese Journal of Analytical Chemistry 2014;(8):1144-1149
A new method for O-ethyl S-[2-( diisopropylamino) ethyl] methylphosphonothiolate ( VX) , sarin detection and its kinetic analysis based on piezoresistive microcantilever aptasensor was developed, where VX, sarin aptamers were immobilized on the microcantilever surface by biotin-avidin binding system. A linear relationship between the response voltage and the concentration of VX in the range of 2-60μg/L was obtained. The linear regression equation was △Ue=0. 886C-1. 039 (n=5, R=0. 984, p<0. 001) and the detection limit was 2μg/L ( S/N≥3 ) . A linear relationship between the response voltage and the concentration of sarin in the range of 10-60 μg/L was obtained, the linear regression equation was △Ue=0. 716C-2. 304 ( n=5, R=0 . 996 , p<0 . 001 ) and the detection limit was 10 μg/L ( S/N≥3 ) . The sensor showed no response for O-butyl methylphosphonochloridate, a structural analog of VX and sarin, which indicated high specificity and good anti-interference ability. On this basis, a reaction kinetic model based on receptor-ligand binding and the relationship with output voltage change was established. Response voltage (△Ue ) and response time( t0 ) were obtained from the fitting equation on different concentrations of VX, sarin fitted well with the measured values.