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Author ORCID Identifier
https://orcid.org/0000-0002-6734-4436
AccessType
Campus-Only Access for Five (5) Years
Document Type
dissertation
Degree Name
Doctor of Philosophy (PhD)
Degree Program
Chemical Engineering
Year Degree Awarded
2024
Month Degree Awarded
February
First Advisor
Sarah L. Perry
Subject Categories
Biochemical and Biomolecular Engineering | Complex Fluids
Abstract
Proteins play a pivotal role in the vitality of living organisms, orchestrating a diverse array of essential functions. Unravelling these functions requires the determination of their three-dimensional structures. The foundational step in the process of structure determination is the formation of protein crystals. In this pursuit, we have engineered microfluidic devices capable of on-chip protein crystallization. Notably, our design incorporates advanced fluid handling systems, realized through both centrifugal and electrically actuated valves, allowing us to depart from conventional pressure-driven systems that are typically associated with cumbersome equipment like pneumatic controllers and nitrogen cylinders. Subsequent to crystallization, the next critical phase involves X-ray crystallography—a technique wherein proteins are subjected to X-ray beams, and the resulting diffractions are meticulously analysed to unveil the underlying structure. To facilitate this, our ultra-thin microfluidic devices are effectively transparent to X-rays, enabling seamless in-situ structure determination. This pivotal innovation eliminates manual intervention in the handling of protein crystals, streamlining the process. Traditionally, X-ray crystallography has been executed under cryogenic conditions to stabilize protein crystals cultivated in well plates. However, this approach restricts our ability to observe proteins in their native physiological conditions. To transcend these limitations, we are actively engaged in two parallel endeavours. Firstly, we have developed scalable devices using roll-to-roll technologies that can be used for data collection at ambient temperatures, and are compatible with robotic handling for seamless automation. Concurrently, we have also developed graphene-based platforms, affording us a unique vantage point to study crystal structures under physiological conditions spanning from 4°C to 70°C. Together, these advances represent enabling technologies that are driven by an engineering approach to solve problems at the cutting edge of structural biology.
DOI
https://doi.org/10.7275/36454131
Recommended Citation
Saha, Sarthak, "ADVANCED MICROFLUIDICS PLATFORMS FOR PROTEIN CRYSTALLIZATION AND X-RAY CRYSTALLOGRAPHY" (2024). Doctoral Dissertations. 3088.
https://doi.org/10.7275/36454131
https://scholarworks.umass.edu/dissertations_2/3088
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.