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Designing Cell-Free Protein Synthesis Systems For Improved Biocatalysis And On-Demand, Cost-Effective Biosensors, Mehran Soltani Najafabadi
Designing Cell-Free Protein Synthesis Systems For Improved Biocatalysis And On-Demand, Cost-Effective Biosensors, Mehran Soltani Najafabadi
Theses and Dissertations
The open nature of Cell-Free Protein Synthesis (CFPS) systems has enabled flexible design, easy manipulation, and novel applications of protein engineering in therapeutic production, biocatalysis, and biosensors. This dissertation reports on three advances in the application of CFPS systems for 1) improving biocatalysis performance in industrial applications by site-specific covalent enzyme immobilization, 2) expressing and optimizing a difficult to express a mammalian protein in bacterial-based CFPS systems and its application for cost-effective, on-demand biosensors compatible with human body fluids, and 3) streamlining the procedure of an E. coli extract with built-in compatibility with human body fluid biosensors. Site-specific covalent immobilization …
Engineering Cell-Free Protein Expression Systems For Biotherapeutics And Biosensing, John Porter Hunt
Engineering Cell-Free Protein Expression Systems For Biotherapeutics And Biosensing, John Porter Hunt
Theses and Dissertations
Therapeutic proteins have become a cornerstone of modern medicine since the FDA approval of recombinant human insulin in 1982. Likewise, biosensors transform chemical detection and disease diagnostics by identifying biomarkers, chemical contaminants, and infective agents. Long-standing methods for creating therapeutics and biosensors employ whole cells such as Escherichia coli (E. coli). Alternatively, cell-free protein synthesis (CFPS) employs the enzymatic reactions necessary for protein production and biosensing within a cell, but in an engineered reactor environment facilitating unprecedented access to and control over biochemical machinery, preservation by cryodesiccation for portable deployment, and functionality in cytotoxic applications. This dissertation reports advances in …