Alpha-oxo semicarbazone peptide or oligodeoxynucleotide microarrays.
We describe in this paper the preparation and characterization of semicarbazide glass slides and their use for the fabrication of microarrays using site-specific alpha-oxo semicarbazone ligation. The functional density and homogeneity of the semicarbazide glass slides were optimized by analyzing the reactivity of the layer toward a synthetic glyoxylyl fluorescent probe. Oligonucleotide microarrays were prepared by site-specific immobilization of glyoxylyl oligodeoxynucleotides. The slides were directly used in the hybridization assays using fluorescence detection and displayed a significant gain in sensibility as compared to the aldehyde glass slide/amino oligodeoxynucleotide chemistry. Semicarbazide slides were also used for the immobilization of a biotinylated peptide alpha-oxo aldehyde. The peptide microarrays allowed model interaction studies with streptavidin or an anti-biotin antibody.
Olivier
Christophe
C
UMR CNRS 8525, Biological Institute of Lille, 1 rue du Pr Calmette 59021 Lille France.
Hot
David
D
Huot
Ludovic
L
Ollivier
Nathalie
N
El-Mahdi
Ouafâa
O
Gouyette
Catherine
C
Huynh-Dinh
Tam
T
Gras-Masse
Hélène
H
Lemoine
Yves
Y
Melnyk
Oleg
O
D000447:Aldehydes / Q000737:chemistry; D000906:Antibodies / Q000737:chemistry; D001710:Biotin / Q000737:chemistry / Q000276:immunology; D002021:Buffers; D005456:Fluorescent Dyes; D006868:Hydrolysis; D017403:In Situ Hybridization; D007202:Indicators and Reagents; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000737:chemistry* / Q000235:genetics; D010455:Peptides / Q000737:chemistry*; D011786:Quality Control; D012663:Semicarbazides / Q000737:chemistry*; D012821:Silanes / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry
D000447:Aldehydes; D000906:Antibodies; D002021:Buffers; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D009841:Oligonucleotides; D010455:Peptides; D012663:Semicarbazides; D012821:Silanes; D001710:Biotin; D019809:Streptavidin