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Detection of molecular interactions is critical for understanding many biological processes, for detecting disease biomarkers, and for screening drug candidates. Fluorescence-based approach can be problematic, especially when applied to the detection of small molecules. Various label-free techniques, such as surface plasmon resonance technique are sensitive to mass, making it extremely

Detection of molecular interactions is critical for understanding many biological processes, for detecting disease biomarkers, and for screening drug candidates. Fluorescence-based approach can be problematic, especially when applied to the detection of small molecules. Various label-free techniques, such as surface plasmon resonance technique are sensitive to mass, making it extremely challenging to detect small molecules. In this thesis, novel detection methods for molecular interactions are described.

First, a simple detection paradigm based on reflectance interferometry is developed. This method is simple, low cost and can be easily applied for protein array detection.

Second, a label-free charge sensitive optical detection (CSOD) technique is developed for detecting of both large and small molecules. The technique is based on that most molecules relevant to biomedical research and applications are charged or partially charged. An optical fiber is dipped into the well of a microplate. It detects the surface charge of the fiber, which does not decrease with the size (mass) of the molecule, making it particularly attractive for studying small molecules.

Third, a method for mechanically amplification detection of molecular interactions (MADMI) is developed. It provides quantitative analysis of small molecules interaction with membrane proteins in intact cells. The interactions are monitored by detecting a mechanical deformation in the membrane induced by the molecular interactions. With this novel method small molecules and membrane proteins interaction in the intact cells can be detected. This new paradigm provides mechanical amplification of small interaction signals, allowing us to measure the binding kinetics of both large and small molecules with membrane proteins, and to analyze heterogeneous nature of the binding kinetics between different cells, and different regions of a single cell.

Last, by tracking the cell membrane edge deformation, binding caused downstream event – granule secretory has been measured. This method focuses on the plasma membrane change when granules fuse with the cell. The fusion of granules increases the plasma membrane area and thus the cell edge expands. The expansion is localized at the vesicle release location. Granule size was calculated based on measured edge expansion. The membrane deformation due to the granule release is real-time monitored by this method.
ContributorsGuan, Yan (Author) / Tao, Nongjian (Thesis advisor) / LaBaer, Joshua (Committee member) / Goryll, Michael (Committee member) / Wang, Shaopeng (Committee member) / Arizona State University (Publisher)
Created2015
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Wilhelm Pfeffer published his book Osmotische Untersuchungen: Studien Zur Zellmechanik (Osmotic Investigations: Studies on Cell Mechanics) in 1877 during his time as a professor of botany at the University of Basel in Basel, Switzerland. Gordon R. Kepner and Eduard J. Stadelmann translated the book into English in 1985. Verlag von

Wilhelm Pfeffer published his book Osmotische Untersuchungen: Studien Zur Zellmechanik (Osmotic Investigations: Studies on Cell Mechanics) in 1877 during his time as a professor of botany at the University of Basel in Basel, Switzerland. Gordon R. Kepner and Eduard J. Stadelmann translated the book into English in 1985. Verlag von Wilhelm Engelmann in Leipzig, Germany, published the original book in German in 1877 and Van Nostrand Reinhold Company in New York, New York, published the English version in 1985. The book focuses on the cell mechanics of osmotic processes to explain why high pressure exists in plant cells. The book also provides one of the earliest detailed descriptions of the Pfeffer Cell, a devise Pfeffer had created to model and study osmosis in plant cells. The model helped Pfeffer propose theories for how osmosis affected metabolism, growth, and development of plant cells.

Created2017-05-09