Activity Study of Self-Assembled Proteins on Nanoscale Diblock Copolymer Templates

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• 作者：Nitin Kumar ; Omkar Parajuli ; Adam Dorfman ; Dylan Kipp ; Jong-in Hahm
• 刊名：Langmuir
• 出版年：2007
• 出版时间：July 3, 2007
• 年：2007
• 卷：23
• 期：14
• 页码：7416 - 7422
• 全文大小：592K
• 年卷期：v.23,no.14(July 3, 2007)
• ISSN：1520-5827

文摘

Novel methods for affixing functional proteins on surfaces with high areal density have the potential to promotebasic biological research as well as various bioarray applications. The use of polymeric templates under carefullybalanced thermodynamic conditions enables spontaneous, self-assembled protein immobilization on surfaces withspatial control on the nanometer scale. To assess the full potential of such nanometer-scale protein platforms inbiosensing applications, we report for the first time the biological activity of proteins on diblock copolymer platforms.We utilized horseradish peroxidase, mushroom tyrosinase, enhanced green fluorescent protein, bovine immunoglobulinG, fluorescein isothiocyanate conjugated anti-bovine IgG, and protein G as model systems in our protein activitystudies. When specific catalytic functions of HRP and MT, immobilized on selective domains of microphase-separatedPS-b-PMMA, are evaluated over a long period of time, these enzymes retain their catalytic activity and stability forwell over 3 months. By performing confocal fluorescence measurements of self-fluorescing proteins and interactingprotein/protein systems, we have also demonstrated that the binding behavior of these proteins is unaffected by surfaceimmobilization onto PS-b-PMMA diblock copolymer microdomains. Our polymer platforms provide highly periodic,high-density, functional, stable surface-bound proteins with spatial control on the nanometer scale. Therefore, ourdiblock copolymer-guided protein assembly method can be extremely beneficial for high-throughput proteomicapplications.