Olivier Christophe, Hot David, Huot Ludovic, Ollivier Nathalie, El-Mahdi Ouafâa, Gouyette Catherine, Huynh-Dinh Tam, Gras-Masse Hélène, Lemoine Yves, Melnyk Oleg
UMR CNRS 8525, Biological Institute of Lille, 1 rue du Pr Calmette 59021 Lille France.
Bioconjug Chem. 2003 Mar-Apr;14(2):430-9. doi: 10.1021/bc025571q.
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.
我们在本文中描述了氨基脲玻片的制备与表征，以及使用位点特异性α-氧代氨基脲连接法将其用于制造微阵列。通过分析该层对合成的乙醛酰基荧光探针的反应性，优化了氨基脲玻片的功能密度和均匀性。通过位点特异性固定乙醛酰基寡脱氧核苷酸制备了寡核苷酸微阵列。这些玻片直接用于使用荧光检测的杂交分析，与醛玻片/氨基寡脱氧核苷酸化学方法相比，灵敏度有显著提高。氨基脲玻片还用于固定生物素化的肽α-氧代醛。该肽微阵列可用于与链霉亲和素或抗生物素抗体进行模型相互作用研究。
