Cell-free Protein Engineering Research

The Bundy Biotechnology research lab studies the expression and engineering of proteins for biocatalysis, vaccination, drug-delivery, and medical diagnostic applications.


Cell-free Synthetic Biology for Protein Engineering

What is Cell-free Synthetic Biology?

Proteins—polymers of amino acids—are a major class of biomolecules whose myriad functions facilitate many crucial biological processes. Accordingly, human control over these biological processes depends upon the ability to study, produce, and modify proteins. One innovative tool for accomplishing these aims is cell-free protein synthesis (CFPS). This method, rather than using living cells to make protein, simply extracts the cells’ natural protein-making machinery and then uses it to produce protein in vitro.

Engineering Within a Cell-free System

Because living cells are no longer involved, scientists can freely adapt and engineer the protein production environment in ways not otherwise possible. This increased accessibility enables cutting-edge research on current topics such as unnatural amino acid insertion, virus-like particle production, enzyme immobilization, and high-throughput assays.

These exciting areas of study have the potential to contribute to biotherapeutics, recombinant enzyme development, cancer research, and more (see image to right).


 Biocatalysis Immobilization

What are Biocatalysts?
Biocatalysts are enzymes that enable chemical reactions to occur faster and with less energy.  Unlike traditional chemical catalysts, biocatalysts are biodegradable, function at low pressures and temperatures, and produce negligible side products. Biocatalysts already play an important role in making drugs, detergents, food products, fuel etc.
Our Research and Biocatalystsbiocatalysis Picture

In many industrial applications, biocatalysts are gradually lost as they “wash away” with product streams. Our lab seeks to immobilize biocatalysts using unnatural amino acids in order to preserve their retention and activity. Unnatural amino acids can act as unique targets for immobilization, increasing our control over the procedure. The process of incorporating unnatural amino acids into biocatalysts can be efficiently carried out in cell-free systems due to their open nature.

Research Specifics
Breakthroughs in enzyme immobilization have enabled increased enzyme recovery and reusability, leading to significant decreases in the cost of enzyme use and fueling biocatalysis growth. However, current enzyme immobilization techniques suffer from leaching, enzyme stability, and recoverability and reusability issues. Moreover, these techniques lack the ability to control the orientation of the immobilized enzymes. To determine the impact of orientation on covalently immobilized enzyme activity and stability, we apply our PRECISE (Protein Residue-Explicit Covalent Immobilization for Stability Enhancement) system to a model enzyme, T4 lysozyme. The PRECISE system uses non-canonical amino acid incorporation and the Huisgen 1,3-dipolar cycloaddition “click” reaction to enable directed enzyme immobilization at rationally chosen residues throughout an enzyme. Unlike previous site-specific systems, the PRECISE system is a truly covalent immobilization method. Utilizing this system, enzymes immobilized at proximate and distant locations from the active site were tested for activity and stability under denaturing conditions.


Virus-like Particles & Cancer Theraputics

The engineering of virus-like particles for highly specific imaging agents, drug delivery vehicles, and vaccine antigens is another major thrust of the Bundy Lab. Separately, the engineering of more effective cancer Therapeutics has been initiated as a new research project.

Icosahedral virus like particle (VLP) is a hollow spherical protein container. Hundreds of identical monomer self-assemble to become a microcompartment. VLPs are genetically modifiable. This property  enables us to purify them, encapsidate guest molecules, or decorate the exterior surface with ligands, dye, etc. Currently many biotechnologists utilize VLPs to make drug delivering vehicles, bioimaging materials, vaccines, biochemical reaction vessels.


Dr. Bundy Shares About Biocatalysis

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Dr. Bundy’s research team would like to thank the following for providing funding for our research.

DARPA (Department of Defense)Funding_logo2

National Science Foundation (CBET Division)

National Pork Board (Foreign Animal Disease)

NASA (Rocky Mountain Space Grant Consortium)

BYU (ORCA/Graduate Studies)