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Issue 8, 2011
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Characterization of protein immobilization on nanoporous gold using atomic force microscopy and scanning electron microscopy

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Abstract

Nanoporous gold (NPG), made by dealloying low carat gold alloys, is a relatively new nanomaterial finding application in catalysis, sensing, and as a support for biomolecules. NPG has attracted considerable interest due to its open bicontinuous structure, high surface-to-volume ratio, tunable porosity, chemical stability and biocompatibility. NPG also has the attractive feature of being able to be modified by self-assembled monolayers. Here we use scanning electron microscopy (SEM) and atomic force microscopy (AFM) to characterize a highly efficient approach for protein immobilization on NPG using N-hydroxysuccinimide (NHS) ester functionalized self-assembled monolayers on NPG with pore sizes in the range of tens of nanometres. Comparison of coupling under static versus flow conditions suggests that BSA (Bovine Serum Albumin) and IgG (Immunoglobulin G) can only be immobilized onto the interior surfaces of free standing NPG monoliths with good coverage under flow conditions. AFM is used to examine protein coverage on both the exterior and interior of protein modified NPG. Access to the interior surface of NPG for AFM imaging is achieved using a special procedure for cleaving NPG. AFM is also used to examine BSA immobilized on rough gold surfaces as a comparative study. In principle, the general approach described should be applicable to many enzymes, proteins and protein complexes since both pore sizes and functional groups present on the NPG surfaces are controllable.

Graphical abstract: Characterization of protein immobilization on nanoporous gold using atomic force microscopy and scanning electron microscopy

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Publication details

The article was received on 27 Apr 2011, accepted on 13 Jun 2011 and first published on 13 Jul 2011


Article type: Paper
DOI: 10.1039/C1NR10427F
Citation: Nanoscale, 2011,3, 3395-3407
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    Characterization of protein immobilization on nanoporous gold using atomic force microscopy and scanning electron microscopy

    Y. H. Tan, J. R. Schallom, N. V. Ganesh, K. Fujikawa, A. V. Demchenko and K. J. Stine, Nanoscale, 2011, 3, 3395
    DOI: 10.1039/C1NR10427F

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