Electrical Readout of Protein Microarrays on Regular Glass Slides


AU: 
Bonilla, D; Mallen, M; de la Rica, R; Fernandez-Sanchez, C; Baldi, A
SO: 
ANALYTICAL CHEMISTRY
Autores del GTQ: 
VL: 
83
IS: 
5
BP: 
1726
EP: 
1731
PY: 
2011
SN: 
0003-2700
Impact: 
5.87
Año IF: 
2010
Resumen: 

A new approach for the electrical readout of microarrays prepared on regular glass slides, using an array of impedimetric transducers (interdigitated electrodes, IDEs) is presented in this work. Impedance detection relies on the use of a urease-labeled immunoassay scheme. Urease is able to produce an increase in conductivity by hydrolysis of the urea substrate, which is measured with the IDEs and directly related to the amount of target analyte. Unlike previous electrical microarrays, the assay does not take place on top of the transducers but on a regular glass slide, which may enable the development of compact multiplexed analytical systems with lower cost per assay. A droplet of solution with the enzymatic substrate is deposited on each transducer of the array, and the microarray is positioned at a short distance (300 urn) so that each droplet wets one transducer and one spot of the microarray. This procedure allows reusing the transducer array for readout of a virtually unlimited number of microarrays. A microarray based on an immunoassay for the detection of a mouse generic protein in a concentration range from 0.03 to 30 mu g mL(-1) was carried out to assess the performance of the electrical readout approach. A sigmoid response with a limit of detection of 0.1 mu g mL(-1) and a dynamic range of 1 order of magnitude was obtained. A comparative study was also carried out with two well established analytical procedures. First, the urease-based immunoassay was tested in a 96 well microtiter plate using phenol red pH indicator and absorbance detection. Second, the microarray was carried out using the same target protein concentration range but applying a Cy3 label and fluorescence detection. Both assays allowed for the validation of the performance of the presented electrical readout system.

DT: 
Article en revistes indexades
Percentil: 
HIGH
Año Percentil: 
2010
Categoría: 
CHEMISTRY, ANALYTICAL