2023 197 E1

A Low-Cost and Reversible Protein Purification Method Based on Optogentic Tools

By: Zachary Pope, Evan Forrest

Department: Biology, Chemistry & Biochemistry 

Faculty Advisor: Dr. Raymond Esquerra

Many research fields and biotechnology industries require fast and cost effective protein purification. To address this challenge, we developed a protein purification strategy based on optogenetic tools that specifically use the Phytochrome B and PIF1 photoreceptors. This method requires binding a light sensitive Phytochrome B to a solid agarose resin and using PIF1 as a protein tag, enabling the binding and release of proteins in response to red and far red light. We demonstrate the effectiveness of this reversible and low cost method by purifying myoglobin and maltose binding protein, and show that binding capacity and purification efficiency are comparable to other biochemical chromatography media. In summary, this PIF-tag system has great potential for achieving fast and affordable protein purification in various applications.