Disposable screen-printed chemiluminescent biochips for the simultaneous determination of four point-of-care relevant proteins.
A screen-printed (SP) microarray is presented as a platform for the achievement of multiparametric biochips. The SP platform is composed of eight (0.28-mm(2)) working electrodes modified with electroaddressed protein A-aryl diazonium adducts. The electrode surfaces are then used as an affinity immobilisation support for the orientated binding of capture monoclonal antibodies, having specificity against four different point-of-care related proteins (myoglobin, cardiac troponin I, C-reactive protein and brain natriuretic peptide). The immobilised capture antibodies are involved in sandwich assays of the four proteins together with biotinylated detection antibodies and peroxidase-labelled streptavidin in order to permit a chemiluminescent imaging of the SP platform and a sensitive detection of the assayed proteins. The performances of the system in pure buffered solutions, using a 25-min assay duration, were characterised by dynamic ranges of 0.5-50, 0.1-120, 0.2-20 and 0.67-67 microg/L for C-reactive protein, myoglobin, cardiac troponin I and brain natriuretic peptide, respectively. The four different assays were also validated in spiked 40-times-diluted human sera, using LowCross buffer, and were shown to work simultaneously in this complex medium.
Marquette
C A
CA
Laboratoire de Génie Enzymatique et Biomoléculaire, Université Lyon 1-CNRS 5246 ICBMS, Bât CPE, 43 bd du 11 novembre 1918, 69622, Villeurbanne Cedex, France. christophe.marquette@univ-lyon1.fr
Bouteille
F
F
Corgier
B P
BP
Degiuli
A
A
Blum
L J
LJ
D004566:Electrodes; D049449:Luminescence; D040081:Protein Array Analysis; D011506:Proteins / Q000737:chemistry*
D011506:Proteins