Atomic Force Microscopy Investigation of the Morphology and the Biological Activity of Protein-Modified Surfaces for Bio- and Immunosensors

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• 作者：Francesca Cecchet ; Anne-Sophie Duwez ; Sabine Gabriel ; Christine Jé ; r&ocirc ; me ; Robert Jé ; r&ocirc ; me ; Karine Glinel ; Sophie Demoustier-Champagne ; Alain M. Jonas ; Bernard Nysten
• 刊名：Analytical Chemistry
• 出版年：2007
• 出版时间：September 1, 2007
• 年：2007
• 卷：79
• 期：17
• 页码：6488 - 6495
• 全文大小：790K
• 年卷期：v.79,no.17(September 1, 2007)
• ISSN：1520-6882

文摘

With the purpose of developing biosensors, the reliableproof of the biological activity of two new sensor systemswas obtained by atomic force microscopy (AFM) in boththe imaging and the single-molecule force spectroscopymodes. Antigens or antibodies of pharmacological interestwere grafted onto self-assembled monolayers of thiols ongold, and AFM imaging demonstrated that the graftingprocess produced homogeneous submonolayers of isolated proteins. The analysis of the morphology of thesurfaces at the different functionalization steps allowedevaluating the protein grafting density and showed thatthe recognition of complementary species present in thesurrounding solution occurred. Single-molecule forcespectroscopy experiments between the sensing surfacesand AFM probes, onto which the complementary specieswere grafted, enabled a direct and rapid test of thebiological activity of the sensors by investigating theinteraction occurring at the level of one single ligand-receptor bond. Ellipsometry and surface plasmon resonance allowed further characterization of the sensorsurfaces and confirmed that the biological recognition tookplace.