Multi-Purpose Hyper-Stable Fluorescent Proteins

Principal Investigator: 

Benjamin Campbell

Background & Unmet Need

  • Fluorescent proteins (FPs) are used for a broad range of biology research applications, including as reporters of gene activity and to visualize proteins in live cells
  • Most FPs lack the ability to withstand the adverse conditions in the cell that rapidly denature biological structures, including superfolder GFP
  • Modern microscopy techniques such as CLEM and ExM, place even greater demands on FPs due to secondary fixation and staining chemicals such as osmium tetroxide that degrade protein functionality
  • Most widely used FPs are susceptible to misfolding due to exposure to higher temperatures for a greater lengths of time and photo bleach quickly
  • Unmet Need: There is a need for stable FPs that can withstand chaotropic conditions and may be used for various biochemical applications

Technology Overview

  • The Technology: Development of extremely stable FPs that withstand chaotropic conditions
  • The engineered FPs Hyperfolder YFP (hfYFP) and LSSmGFP successfully survive CLEM sample preparation and ExM techniques
  • hfYFP contains no cysteines, is chloride insensitive, and tolerates aldehyde and osmium tetroxide fixation better than common FPs
  • The robust nature of hfYFP and LSSmGFP make them ideal FPs for the development of biosensors
  • PoC Data: The chemical stability of hfYFP was employed to devise a fluorescence-assisted protein purification strategy, enabling all steps of denaturing affinity chromatography to be visualized using UV or blue light

Technology Applications

  • Fluorescence-assisted protein purification
  • Live cell imaging and the development of improved biosensors
  • Stable FPs for use with CLEM and ExM techniques

Technology Advantages

  • Compatible with osmium tetroxide and aldehyde fixation protocols
  • Chloride insensitive
  • Improved acid resistance
  • Greater thermodynamic stability and faster refolding than existing FPs

Comparison of different fluorescent proteins

Intellectual Property

Patents

  • PCT Application Filed WO2024015362A1: "Chemically stable fluorescent proteins for advanced microscopy"

Cornell Reference

  • 10194

Contact Information

Dr. Jeff James

For additional information please contact

Jeffrey James
Associate Director, Business Development and Licensing
Phone: (646) 962-4194
Email: jaj268@cornell.edu