| This dissertation describes two projects aimed at developing: (1) a pair of nanofabrication and analytical tools that allow one to deliberately control biomolecular surface architecture on the 1--100 nm length, and (2) a novel detection method for measuring extraordinarily small numbers of Human immunodeficiency virus type-1 (HIV-1) marker proteins in complex environments. The first project approach deals with the development of new chemistry and methods for fabricating nanoarrays comprised of different proteins and the study of the fundamental interactions between such arrays and biological structures (cells, complementary proteins, and viruses). These nanoarrays have provided avenues to new and powerful ways of detecting protein markers, studying cell-surface interactions, and organizing biological entities such as viruses into higher ordered architectures. In chapter 2, nanoarrays of retronectin are described, and their ability to establish focal adhesion between cells and surfaces, even when the feature sizes are approximately 200 nm, is demonstrated. This was the first time sub-micrometer patterns have been utilized to effect such a process, and it shows that receptor ligand-interactions are a nanoscale rather than microscale phenomenon, even when ligand clustering takes place. Chapter 3 describes the use of nanoarrays to organize Cowpea mosaic virus (CPMV) in a chemoselective manner. This work shows that two different virus particles can be differentiated with nanopatterns based upon surface architecture and virus capsid proteins. Finally, in Chapter 4, the chemistry and Dip-Pen Nanolithography (DPN) methodology for directly patterning a wide variety of surfaces, including Au, SiOx, and NiOx, with intact and bioactive protein structures is demonstrated.; The second project pertains to the development of two new protein detection methods based upon nanoarrays and nanoparticle probes. Both have focused on HIV-1 proteins as targets. In chapter 5, it is demonstrated that nanoarrays can be used in a probeless detection method that involves the screening of an entire array with an Atomic Force Microscope (AFM). In chapter 6, a Bio-Barcode Amplification (BCA) assay for detecting a wide variety of HIV protein markers is reported.; Finally, in chapter 7, a novel approach to separating target protein from complex solutions containing multiple proteins is reported. This method utilizes magnetic multicomponent gold-nickel nanorods as a separation agent. (Abstract shortened by UMI.)... |
