Novel Method to Monitor the Viral Proteome Throughout Infection for Use in Anti-Viral Drug Discovery and Vaccine Development

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Novel Method to Monitor the Viral Proteome Throughout Infection for Use in Anti-Viral Drug Discovery and Vaccine Development

Princeton Docket # 20-3633

Researchers in the department of molecular biology at Princeton University have developed novel assays that use a targeted mass spectrometry approach to monitor viral proteins from all distinct temporal classes of a virus replication cycle within host cells. The replication of a herpesvirus in an infected cell and the spread of infection to neighboring cells relies on a finely controlled lifecycle with a temporally tuned cascade of viral gene expression. In order to effectively identify potential virus modulatory compounds, as well as gain an understanding of their impact on specific stages of a viral infection, we have developed a novel assay to monitor viral proteins from herpesviruses. This assay offers the accurate detection and quantification of viral proteins from all distinct temporal classes of viral replication. We have designed three assays for the specific detection of three herpesviruses: the α-herpesvirus herpes simplex virus 1 (HSV1), the β-herpesvirus human cytomegalovirus (HCMV), and the γ-herpesvirus Kaposi’s sarcoma herpesvirus (KSHV). These assays can be utilized in combination with drug treatments, genetic modifications, or other perturbations to assess the impact of the intervention on viral protein production. Given the temporal nature of herpesvirus infection, the acquired protein abundance measurements provide information regarding the stage of infection that is affected, the specific viral proteins that are impacted, as well as an additional mechanistic understanding of how a given compound or other perturbation impacts viral replication.

Drug companies working on antiviral therapies or vaccine development can use these assays either as a primary screen for initial anti- or pro-viral activity identification or as a secondary screen to provide additional information on compounds that have been determined to impact infection through titer assays or other endpoint screening techniques. Compared to the current standard approach of a broad chemical compound screen with a reduction in viral titer as the endpoint, the proposed method is more efficient and accurate. It helps shortcut the mechanistic investigations by immediately focusing future work on a subset of the infection process. Other competing technologies used for antiviral compound discovery include mRNA sequencing, gene microarray, and western blotting analysis. In comparison, the proposed assays exceed these methods both in detection specificity and quantification.  


  • Primary screen for initial anti- or pro-viral activity identification of small molecules
  • Secondary screen for additional information on compounds (such as functions, impacts, and subcellular localizations)
  • Secondary screen for confirmation and additional information on cellular factor anti- or pro-viral activity
  • Facilitate anti-viral therapy and vaccine development


  • High specificity of detection
  • Accuracy of quantification
  • Time efficiency

Stage of Development

A working prototype was developed, successfully measuring the progression of infection for three herpesviruses that are major human pathogens: Herpes Simplex Virus 1 (HSV1), Human Cytomegalovirus (HCMV), and Kaposi’s Sarcoma Herpesvirus (KSHV).


Ileana Cristea graduated with her Ph.D. in 2002 from the Michael Barber Center for Mass Spectrometry, UMIST (University of Manchester Institute of Science and Technology), UK, performed her postdoctoral research at Rockefeller University, and is a Professor in the Department of Molecular Biology at Princeton University. Her group, the Cristea Laboratory, is poised at the interface of virology and proteomics, and aims to understand the mechanisms of cellular host defense against infection with human viruses.

Joel Federspiel graduated with his Ph.D. in Biochemistry from Vanderbilt University in 2016 and pursued his postdoctoral work in the Cristea Laboratory at Princeton University (2016-19). He uses mass spectrometry to investigate how viral proteins suppress or hijack host proteins in order to identify potential novel antiviral targets.

Intellectual Property Status

Patent protection is pending.
Industry collaborators are sought to further develop and commercialize this technology.


Laurie Tzodikov
Princeton University Office of Technology Licensing • (609) 258-7256•

Isla Xi Han
Princeton University Office of Technology Licensing •

Patent Information:
For Information, Contact:
Prabhpreet Gill
Licensing Associate
Princeton University
Ileana Cristea
Joel Federspiel
Michelle Kennedy