Split-Supercharged GFP: A Robust Platform to Detect Protein-Protein Interactions

Figure 1. PC-3 cells following treatment with 100 nM split-spGFP prostate cancer cell-binding peptide fusions.

Available for Licensing

IP Status

US Utility Patent: US 9400249


Brian McNaughton
Brett Blakely
Alex Chapman

At A Glance

Split-supercharged GFP reassembly is an operationally simple and robust technique to identify and study protein-protein, peptide-protein, and peptide-peptide interactions (PPIs) in living cells. The susceptibility of other split-GFP reassembly systems to undergo rapid and irreversible aggregation places dramatic limitations on their use. In contrast, split-supercharged GFP is resistant to aggregation, and undergoes rapid (<3 hours) and efficient interaction-dependent reassembly – dramatically simplifying the identification of interacting pairs in E. coli. In contrast to other split-GFP systems, split-supercharged GFP works well at physiological temperature. The efficiency, robustness, and simplicity of split-supercharged GFP reassembly make this reporter system ideal for identifying and studying interactions involving proteins and/or peptides in living cells, and may be particularly useful for identifying or studying interactions involving proteins or peptides that are themselves susceptible to aggregation.

This particular technology has been distributed to academic collaborators via AddGene over 400 times in the past 2 years, showing remarkable usage.  With the backing of a life science company, this technology would be utilized worldwide.


Licensing Director

Steve Foster

Reference No.: 12-100

  • Rapid detection of polypeptide interactions inside coli
  • High signal-to-noise
  • Efficient peptide-peptide interaction detection at physiological temperature
  • Detection of interactions involving aggregation-prone polypeptides

Blakely, B.D.; Chapman, A.M.; McNaughton, B.R. Molecular BioSystems 2012, 8, 2036

Chapman, A.M.; McNaughton, B.R. ACS Chemical Biology 2014, 9, 2223

Walker, S.N.; Tennyson, R.L.; Chapman, A.M.; Kennan, A.J.; McNaughton, B.R. ChemBioChem 2015, 16, 2

Last updated: May 2020