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多肽氨基脲玻璃片微阵列:血清学检测研究中的特性及与胺基片的比较

Polypeptide semicarbazide glass slide microarrays: characterization and comparison with amine slides in serodetection studies.

作者信息

Duburcq Xavier, Olivier Christophe, Desmet Rémi, Halasa Matej, Carion Olivier, Grandidier Bruno, Heim Thomas, Stiévenard Didier, Auriault Claude, Melnyk Oleg

机构信息

UMR CNRS 8527, Biological Institute of Lille, 1 Rue du Pr Calmette 59021 Lille, France.

出版信息

Bioconjug Chem. 2004 Mar-Apr;15(2):317-25. doi: 10.1021/bc034118r.

Abstract

We have described in the accompanying article the preparation of peptide-protein semicarbazide microarrays and their use for the simultaneous serodetection of antibodies directed against different pathogens. Here, we present a comparative study between semicarbazide and amine glass slides in an immunofluorescent serodetection assay using HIV (Gp120, Gp41), HCV (mix-HCV, core, NS3, and NS4), and HBV (HBs) recombinant antigens. Amine and semicarbazide surfaces displayed the same sensitivity for antibodies detection just after printing. However, the reactivity of protein antigens changed rapidly upon aging on amine slides but not on semicarbazide slides. Peptide or protein semicarbazide microarrays were found to be remarkably stable for months. Additional data concerning the characterization of the semicarbazide surface (homogeneity of the slides, chemical stability, contact angle measurements, atomic force microscopy studies, reproducibility of serodetection results) are also presented and discussed.

摘要

我们已在随附文章中描述了肽-蛋白质氨基脲微阵列的制备及其用于同时血清检测针对不同病原体的抗体的用途。在此,我们展示了在使用HIV(Gp120、Gp41)、HCV(混合-HCV、核心、NS3和NS4)以及HBV(HBs)重组抗原的免疫荧光血清检测试验中,氨基脲玻片和胺基玻片之间的比较研究。刚打印后,胺基和氨基脲表面在抗体检测方面表现出相同的灵敏度。然而,蛋白质抗原在胺基玻片上老化时反应性迅速变化,而在氨基脲玻片上则不然。发现肽或蛋白质氨基脲微阵列在数月内都非常稳定。还展示并讨论了有关氨基脲表面特性的其他数据(玻片的均匀性、化学稳定性、接触角测量、原子力显微镜研究、血清检测结果的可重复性)。

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