Micropillar array chips toward new immunodiagnosis.
In this paper, we demonstrate the possibility to use a micropillar array to perform molecular immunodiagnosis. A polydimethylsiloxane (PDMS) microdevice consisting of a rectangular array of micropillars (45 µm in height, 100 × 100 µm square cross section) was used to replace microchannels or gels (polyacrylamide or agarose) to perform electrokinetic separation. This microarray was used to mimic highly diluted gel and to maintain electrolyte within the pillar zone by capillary effect. The electrolyte composition (glycerol and agarose content) was investigated in order to improve protein separation by isoelectric focusing (IEF). The influence of glycerol on focusing time and on the different evaporative contributions was further evaluated. In order to perform an immunodiagnostic of milk allergy, different surface treatments were optimized to prevent milk allergen adsorption on PDMS surface. Poly(dimethylacrylamide)-co-allyl glycidyl ether (PDMA-AGE) as well as gelatin led to a satisfactory signal to noise ratio. Finally the possibility to perform protein mixture separation using this micropillar array chip followed by immunoblotting was demonstrated by using the serum from an allergic individual, confirming the great potential of this analytical platform in the field of immunodiagnosis.
Li
Hong-Yi
HY
Physicochimie des Electrolytes, Colloïdes et Sciences Analytiques (PECSA) UMR 7195, 10 rue Vauquelin, 75231 Paris cedex 05, France.
Dauriac
Virginie
V
Thibert
Valerie
V
Senechal
Hélène
H
Peltre
Gabriel
G
Zhang
Xin-Xiang
XX
Descroix
Stéphanie
S
D003124:Colorimetry / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007118:Immunoassay / Q000295:instrumentation*; D046228:Microarray Analysis / Q000295:instrumentation*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D011933:Reagent Kits, Diagnostic
D011933:Reagent Kits, Diagnostic