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Stabilization of Near-Infrared Fluorescent Proteins by Packaging in Virus-like Particles
Soumen Das1, Liangjun Zhao1, Stephen N Crooke1
1School of Chemistry and Biochemistry, Georgia Institute of Technology, 901 Atlantic Drive, Atlanta, Georgia 30306, United States.
Biomacromolecules
|May 23, 2020
Summary
Near-infrared fluorescent virus-like particles (VLPs) were created using biliverdin-dependent proteins. These engineered VLPs show enhanced stability and are easily detectable for in vivo imaging applications.
Area of Science:
- Biotechnology
- Fluorescent Protein Engineering
- Nanomedicine
Background:
- Virus-like particles (VLPs) offer a versatile platform for biomedical applications due to their inherent biocompatibility and self-assembly properties.
- Near-infrared (NIR) fluorescent probes are crucial for deep-tissue imaging, but their stability and efficient production remain challenges.
- Biliverdin-dependent fluorescent proteins provide a unique spectral range, but their utility is often limited by stability issues.
Purpose of the Study:
- To develop high-yield production of NIR fluorescent Qβ VLPs.
- To enhance the stability and imaging capabilities of biliverdin-dependent fluorescent proteins through encapsulation within VLPs.
- To evaluate the in vivo biodistribution and detectability of these engineered VLPs.
Main Methods:
- Production of monomeric and dimeric biliverdin-dependent fluorescent proteins within the Qβ virus-like particle capsid.
- Simple addition of biliverdin hydrochloride during or after Escherichia coli protein expression for fluorophore maturation.
- Assessment of photochemical properties, stability (heat, chaotrope, proteolysis), and in vivo imaging performance in mice.
Main Results:
- High-yield production of NIR fluorescent Qβ VLPs achieved.
- Encapsidated NIR fluorescent proteins demonstrated identical photochemical properties to non-encapsidated versions.
- Packaged proteins exhibited significantly enhanced stability against heat, denaturation, and proteolysis.
- In vivo imaging in mice revealed primary VLP trafficking to the liver with easy detectability using standard instrumentation.
Conclusions:
- Biliverdin-dependent fluorescent proteins can be efficiently packaged into Qβ VLPs to create stable, NIR-emitting nanoprobes.
- VLP encapsulation significantly improves the robustness of these fluorescent proteins for biological applications.
- The developed NIR fluorescent VLPs are suitable for noninvasive in vivo imaging, with potential applications in diagnostics and research.
