A separation-free amperometric immunosensor for vitellogenin based on screen-printed carbon arrays modified with a conductive polymer.
A disposable amperometric immunosensor was studied for the rapid detection of carp (Carassius auratus) Vitellogenin (Vtg). The sensor was fabricated based on screen-printed carbon arrays (SPCAs) containing eight carbon working and an integrated carbon counter electrodes. To construct the sensor, a conducting polymer (poly-terthiophene carboxylic acid) was electropolymerized on the surface of working electrodes and the polymer-coated SPCAs was characterized by SEM. Horseradish peroxidase (HRP) and a monoclonal antibody (anti-Vtg) specific to carp Vtg were covalently attached onto the polymer modified SPCAs. The immobilization of HRP and anti-Vtg onto the polymer-coated SPCAs was examined using cyclic voltammetry and quartz crystal microbalance studies. In order to detect the amount of Vtg, glucose oxidase (GOx)-labelled Vtg bound to the sensor surface under competition with the Vtg analyte was quantified amperometrically using glucose as a substrate. The performance of the eight sensors in arrays was evaluated by obtaining the calibration plots for Vtg. The sensor arrays exhibit a linear range of the Vtg concentration from 0.25 to 7.8 ng/ml and the detection limit was determined to be 0.09 ng/ml. Furthermore, the performance of the immunosensor for the determination of Vtg was evaluated by a standard addition method performed in fish serum samples.
Darain
Farzana
F
Department of Chemistry and Center for Innovative Bio-physio Sensor Technology, Pusan National University, Pusan 609-735, South Korea.
Park
Doeg Su
DS
Park
Jang-Su
JS
Chang
Seung-Cheol
SC
Shim
Yoon-Bo
YB
D000818:Animals; D015415:Biomarkers / Q000097:blood; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D001774:Blood Chemical Analysis / Q000295:instrumentation / Q000379:methods*; D002244:Carbon / Q000737:chemistry; D002347:Carps / Q000097:blood*; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004553:Electric Conductivity; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes; D005260:Female; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D008297:Male; D008422:Materials Testing; D011108:Polymers / Q000737:chemistry; D013499:Surface Properties; D014819:Vitellogenins / Q000097:blood* / Q000276:immunology; D014874:Water Pollutants, Chemical / Q000097:blood
D015415:Biomarkers; D020099:Coated Materials, Biocompatible; D011108:Polymers; D014819:Vitellogenins; D014874:Water Pollutants, Chemical; D002244:Carbon