Nanoscale protein arrays have shown promise for biological and biomedical applications. Compared to traditional protein arrays, nanoarrays have the potential for higher throughput, better sensitivity, and require less sample volumes. In this thesis, protein nanoarrays were fabricated using a simple and inexpensive "natural lithography" approach. This method allows the fabrication of large-area ordered nanoparticle arrays consisting of metallic dots with tunable diameters down to 10 nm or less.
The nanoparticle arrays are formed by depositing metal through the openings of colloidal monolayer polystyrene sphere masks. After removing the masks, nanoarrays remain and are exposed to further processing. COOH-terminated self-assembled monolayers (SAM) and N-hydroxysuccinimide (NHS) chemistry is used for surface functionalization. These surface modifications covalently attach proteins onto the nanoparticles. A single monolayer of immunoglobulin G (IgG) molecules is successfully attached on the functionalized surfaces and the bioactivity of the protein arrays is tested by attaching anti-IgG molecules, as a standard immunological assay.
Results of fabrication trials and efforts to control nanoparticle size, spacing, and surface adhesion are described. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) images of hexagonal gold nanoarrays consisting of approximately 150 nm particles and 3.5x108-1.5x109 per cm2 array density is shown, depending on the size of colloidal spheres. An increased height of approximately 6 nm characterized via scanning probe methods shows the attachment of a single monolayer of protein molecules to the nanoparticles. This was confirmed with SEM. A similar height increase was detected via AFM showing the attachment of anti-IgG molecules onto IgG functionalized particles. Potential applications of the protein nanoarrays and future work are discussed.
| Identifer | oai:union.ndltd.org:uvic.ca/oai:dspace.library.uvic.ca:1828/2562 |
| Date | 12 April 2010 |
| Creators | Li, Huiyan |
| Contributors | Papadopoulos, Christo |
| Source Sets | University of Victoria |
| Language | English, English |
| Detected Language | English |
| Type | Thesis |
| Rights | Available to the World Wide Web |
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