Conducting elastomer surface texturing: a path to electrode spotting. Application to the biochip production.
A new active support for electro-chemiluminescent biochip preparation has been developed. This material was based on an original material composed of graphite modified polydimethyl siloxane (PDMS). The addressed inclusion of Sepharose beads at the surface of this elastomeric electrode generated interesting local high specific surface. The electrode was characterised by electrochemical (cyclic voltametry, chronoamperomatry) and imaging (scanning electron microscopy (SEM)) methods, and a surface area increase factor of 50 was found, linked to the texturing of the surface generated by the presence of the Sepharose beads. The consequence of this increase was shown to be a jump of the local electrochemical activity which induced a well defined and localised electro-chemiluminescent signal. The new material was used to design biochips based on the electro-chemiluminescent reaction of luminol with enzymatically produced hydrogen peroxide. Thus, when using beads bearing bio-molecules such nucleic acid or human IgG, in conjunction with glucose oxidas-labelled DNA or antibody, sensitive biochips could be obtained with detection limits of 10(11) and 10(10) molecules, respectively. Multi-parameter enzyme-based biochips could also be achieved by locally adsorbing, at the PDMS-graphite surface, either glucose oxidase, lactate oxidase or choline oxidase. Detection limits of 10 microM for lactate and choline and 20 microM for glucose were found, with detection ranging over two decades at least.
Laboratoire de Génie Enzymatique et Biomoléculaire, Université Claude Bernard Lyon 1, UMR 5013 EMB2-CNRS, Bât. CPE, Building 43, 11 November 1918, Villeurbanne 69622, Cedex, France. christophe.marquette@univ-lyon1.fr
Christophe A
CA
Marquette
Loïc J
LJ
Blum
D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D004553:Electric Conductivity; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004566:Electrodes; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods; D008163:Luminescent Measurements / Q000295:instrumentation* / Q000379:methods; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods; D009757:Nylons / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods; D013499:Surface Properties
D020099:Coated Materials, Biocompatible; D004129:Dimethylpolysiloxanes; D009757:Nylons; C085707:poly(dimethylsiloxane)-polyamide copolymer