Engineered Programmable Molecular Scaffolds from Porous Protein Crystals

Fluorescent protein crystals loaded with a macromolecule

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Christopher Snow 

Thaddaus R Huber

Reference No: [techID#]
Licensing Manager

Jessy McGowan

At a Glance

Researchers at Colorado State University have developed a newly patented class of protein crystals that serve as scaffolds to precisely organize diverse guest domains (such as proteins, nucleic acids, nanoparticles, and small molecules) in three-dimensional space.  Unlike other materials, engineered protein crystals are sufficiently organized to allow high-resolution structure determination, yet enable site-specific changes via genetic or chemical modification.  This platform nanotechnology is a key advantage in advanced material engineering where precisely positioned active domains are required.


A key motivation for nano-biotechnology efforts is the creation of designer materials in which the assembly acts to organize functional domains in three dimensions. Crystalline materials are ideal from the validation perspective because X-ray diffraction can elucidate the atomic structure. Relatively little work has focused on engineering protein crystals as scaffolds for nanotechnology, due to the technical challenges of coaxing typical proteins into crystallizing, and the likelihood of disrupting the crystallization process if changes are made to the monomers.


The porous nanocrystals may be hosts for guest macromolecules, including proteins, DNA, active ingredients, or other molecules. Data has been collected regarding the stability and recyclability of the porous crystals to store and release guest molecules in predictable and tunable methods.

Figure 1 (below) illustrates the porous nanocrystal at an elevated temperature increased the reaction kinetics, showing the ability of the nanocrystal to serve as a catalyst for reactions with tunable properties for guest macromolecules

Graph of protein immobilization with the porous nanocrystals

Figure 1. At 45°C, the free enzyme in solution (45C free) had lower rates of product formation than free enzyme at room temperature (20CFree). Immobilized hHRP incubated at elevated temperature had a higher rate of product formation than the same concentration of immobilized enzyme at room temperature. 
Notes: All reactions were performed with 100 μM AmplexRed and 100 μM H2O2.  Resorufin production was monitored under 561 nm wavelength excitation
using a fluorescence plate reader. 
  • Highly stable
  • Controlled loading and unloading
  • Engineered for non-covalent and covalent capture of guest macromolecules
  • Allow for programmed placement within materials
  • Integrated crystals have spatially segregated loading patterns
  • Immobilized enzymes or enzyme pathways (protein zeolites)
  • Host-guest approach to structure determination via x-ray diffraction
  • Advanced delivery sensing or theragnostic materials
  • Transport of therapeutic macromolecules for advanced drug delivery applications
  • Confinement of fluorescent guests for biodegradable and adaptable biosensor