Fluorescent Virus-Like Particles Design

Fluorescent virus-like particles represent a convenient tool for characterization of virus-specific mechanisms and visualizing receptor-ligand interactions. Creative Biolabs provides several approaches to label viral particles, including chemical coupling of fluorophores to the virus surface or genetic fusion of fluorescent proteins to VLPs.

Why Use Fluorescent Virus-like Particles?

Early research of viruses was accomplished using electron microscopy, of which the main disadvantage is that the cells to be analyzed must be immobilized and therefore cannot be applied to living cells, making kinetic studies difficult. The growing variety of available fluorescent dyes and the development of fluorescence microscopy allows direct observation of the interaction between the entire virus and host cells. Moreover, the ability to locate and track viral proteins at the subcellular level lays a solid foundation for the development of real-time single-virus tracking methods within live cells. In addition, fluorescent proteins enable researchers to locate and track viral particles in vivo. Furthermore, the development of flow cytometry has made it possible to directly quantify virus-host interactions.

Several approaches to label viral-like particles have been developed in Creative Biolabs over the last decades, which can generally be divided into chemical coupling of fluorophores to the virus surface or genetic fusion of fluorescent proteins to VLPs.

Methods applied for the labeling of viral particles.

Fig.1 Methods applied for the labeling of viral particles. (Wojta-Stremayr D, 2013)

Chemical Labeling of Viral-like Particles

The chemical compounds can be coupled to the viral particles either covalently or non-covalently. Chemical labeling can also be performed after the virus particles have already assembled.

  • Covalent attachment: fluorophores containing amine-reactive groups.
  • Non-covalent attachment: lipophilic membrane dyes and dyes for the viral genome; biarsenical dyes.

Creative Biolabs provides chemical labeling of VLP using fluorescent labels, including but not limited to:

  • PKH67
  • PKH26
  • DiD/Dil
  • Octadecyl Rhodamine B Chloride (R18)
  • [Ru(phen)2(dppz)]2+
  • Fluorescein
  • Resurfin
  • FITC
  • Rhodamine derivates
  • Alexa Fluor 488
  • Alexa Fluor 594
  • Cy3
  • Cy5
  • CypHer5
  • Quantum dots

Genetic Fusion of Fluorophores to Proteins of VLPs

By fusion of genes encoding viral and fluorescent proteins, a variety of different viruses can be fluorescently labeled, either DNA or RNA, enveloped or non-enveloped. Using this method, each particle carries the genetic information of the fluorescently labeled viral protein, thus each generation of viral progeny will also express the labeled protein. Therefore, genetic labeling allows for the exploration of larger aspects of the viral life cycle as compared to chemical markers that allow for the study of only early events of viral infection. Moreover, simultaneous labeling of several viral proteins with fluorescent proteins that emit light of different wavelengths allows tracking of the fate of such viral proteins after viral infection and during viral assembly.

  • GFP
  • mRFP
  • mCherry
  • YFP
  • CFP

In the process of VLP based vaccines design, development, and manufacturing, working with partners who understand the challenges can dramatically increase efficiency. Creative Biolabs' proven fluorescent VLPs experience can help you overcome the challenges in the VLP vaccines development process.

Reference

  1. Wojta-Stremayr D; et al. Fluorosomes: fluorescent virus-like nanoparticles that represent a convenient tool to visualize receptor-ligand interactions. Sensors (Basel). 2013, 13(7): 8722-49.

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