Protein microarrays on hybrid polymeric thin films prepared by self-assembly of polyelectrolytes for multiple-protein immunoassays
Protein microarrays on hybrid polymeric thin films prepared by self-assembly of polyelectrolytes for multiple-protein immunoassays
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DOI：
10.1002/pmic.200500138
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发表时间：
2006-03-01
期刊：
影响因子：
3.4
通讯作者：
Zhou, JZ
中科院分区：
文献类型：
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作者：
Zhou, XC;Zhou, JZ
在这里，我们报告的发展和表征的蛋白质微阵列的纳米工程3-D纳米薄膜（PET）沉积在载玻片上的连续吸附的聚电解质通过自组装技术。通过静电吸附和三维聚合物膜的多孔结构的截留，将抗体或抗原固定在PET涂覆的载玻片中，从而建立用于平行分析的平台。在PET包被的载玻片上制作抗原和抗体微阵列，并证明了用于多分析物检测的蛋白质微阵列的直接和间接免疫测定。在这些PET包被的载玻片上产生的微阵列具有一致的斑点形态，并提供蛋白质组学分析所需的性能特征。PET膜上的蛋白质微阵列提供低至6 pg/mL的LOD和高达三个数量级的动态范围，这比在醛和聚-L-赖氨酸功能化载玻片上制造的蛋白质微阵列更宽。通过水溶液中的自组装技术构建的PET膜是基于绿色化学的在玻璃表面上产生3-D薄膜涂层的成本有效的方法，并且涂覆的载玻片非常适合于固定许多类型的生物分子，使得可以在这种类型的载玻片上开发各种各样的微阵列格式。
We report here the development and characterization of protein microarrays fabricated on nanoengineered 3-D polyelectrolyte thin films (PET) deposited on glass slide by consecutive adsorption of polyelectrolytes via self-assembly technique. Antibodies or antigens were immobilized in the PET-coated glass slides by electrostatic adsorption and entrapment of porous structure of the 3-D polymer film and thus establishing a platform for parallel analysis. Both antigen and antibody microarrays were fabricated on the PET-coated slides, and direct and indirect immunoassays on protein microarrays for multiple-analyte detection were demonstrated. Microarrays produced on these PET-coated slides have consistent spot morphology and provide performance features needed for proteomic analysis. The protein microarrays on the PET films provide LOD as low as 6 pg/mL and dynamic ranges up to three orders of magnitude, which are wider than the protein microarrays fabricated on aldehyde and poly-L-lysine functionalized slides. The PET films constructed by self-assembly technique in aqueous solution is green chemistry based, cost-effective method to generate 3-D thin film coatings on glass surface, and the coated slide is well suited for immobilizing many types of biological molecules so that a wide variety of microarray formats can be developed on this type of slide.