Electroaddressed immobilization of recombinant HIV-1 P24 capsid protein onto screen-printed arrays for serological testing.
A serological chemiluminescent biochip was designed based on screen-printed electrode arrays composed of nine 1-mm(2) electrodes. Arrays were shown to be produced with good batch-to-batch reproducibility (standard deviations of 4.4 and 12.0% for ferricyanide oxidation potential and current, respectively) and very good reproducibility within a particular array (2.0 and 7.5% standard deviations for the same controls). Electrode arrays were used to electroaddress various bioconjugate structures comprising a recombinant HIV-1 P24 capsid protein (RH24K) in polypyrrole film. Entrapment of RH24K preimmobilized onto maleic anhydride-alt-methyl vinyl ether copolymer was shown to be the more efficient immobilization procedure. This addressed sensing layer enabled the detection of anti-P24 antibodies at a concentration of 3.5 ng/ml through peroxidase-labeled anti-human immunoglobulin G reaction. The biochip was used to perform an HIV-1 serological test in human sera. HIV-1 seropositive and seronegative sera were easily discriminated using serum dilutions greater than 1/10,000.
Marquette
Christophe A
CA
Laboratoire de Génie Enzymatique et Biomoléculaire, EMB2 UMR 5013 CNRS-Université Lyon 1, Bâtiment CPE-43, Bd du 11 Novembre 1918-69622 Villeurbanne, Cedex, France. christophe.marquette@univ-lyon1.fr
Imbert-Laurenceau
Emmanuelle
E
Mallet
Francois
F
Chaix
Carole
C
Mandrand
Bernard
B
Blum
Loïc J
LJ
D000595:Amino Acid Sequence; D004566:Electrodes; D015483:HIV Antibodies / Q000032:analysis* / Q000276:immunology*; D016655:HIV Core Protein p24 / Q000737:chemistry* / Q000276:immunology*; D018023:HIV Seronegativity / Q000276:immunology; D006679:HIV Seropositivity / Q000276:immunology; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007700:Kinetics; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D015203:Reproducibility of Results; D016415:Sequence Alignment; D012698:Serologic Tests / Q000295:instrumentation* / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D015483:HIV Antibodies; D016655:HIV Core Protein p24; D007106:Immune Sera; D011994:Recombinant Proteins