A targeted mass spectrometry platform for high-throughput detection and quantification of herpesvirus infections

Background

Herpesviruses are a family of DNA viruses that are highly prevalent in the global population, with some estimates suggesting that up to 90% of individuals are infected with at least one type. These viruses, which include herpes simplex virus 1 (HSV-1), human cytomegalovirus (HCMV), and Kaposi’s sarcoma-associated herpesvirus (KSHV), can cause a range of diseases from mild cold sores to severe complications in immunocompromised patients and newborns. Accurate detection and monitoring of herpesvirus infections are crucial for effective clinical management, epidemiological studies, and the development of antiviral therapies. The ability to track viral protein expression over the course of infection is particularly important for understanding viral replication dynamics and evaluating the efficacy of potential treatments.

Despite the importance of monitoring herpesvirus infections, current detection methods face significant limitations. Traditional antibody-based assays, such as western blotting and ELISA, often suffer from issues like cross-reactivity, limited specificity, and variability between batches. These methods also lack the capacity for high-throughput multiplexing, making it difficult to simultaneously monitor multiple viral proteins or different stages of infection. Nucleic acid-based techniques, while sensitive, do not provide direct information about viral protein expression, which is essential for assessing active infection and the effects of antiviral drugs. Furthermore, the absence of effective, non-toxic antiviral treatments and vaccines underscores the need for more precise and scalable tools to support both clinical diagnostics and research into herpesvirus biology and therapeutics.

Technology

This technology is a high-throughput assay platform that leverages parallel reaction monitoring (PRM) combined with liquid chromatography tandem mass spectrometry (LC-MS/MS) to detect and quantify herpesvirus proteins directly from infected samples. By utilizing a library of unique, experimentally validated tryptic peptides—each serving as a signature for specific viral proteins—the system can monitor proteins from all stages of viral replication (immediate-early, early, late) across major human herpesviruses such as HSV-1, HCMV, and KSHV. The workflow involves cell lysis, protein digestion, peptide cleanup, and scheduled PRM analysis, with quantification normalized against host protein controls. Heavy isotope-labeled peptide standards are included for precise measurement, and the platform supports multiplexing to cover 30–70% of the viral proteome in a single run. The assay is optimized for automation, scalability, and rapid turnaround, requiring as few as 10,000 cells per sample and generating standardized reports through integrated software.

This solution is differentiated by its direct, protein-level measurement of viral infection, overcoming the limitations of traditional antibody-based assays and nucleic acid detection methods. Unlike conventional approaches, which often suffer from cross-reactivity, limited multiplexing, and indirect measurement of infection status, this platform provides high specificity through signature peptides and robust quantification using mass spectrometry. Its adaptability to widely available instruments, rapid and scalable workflow, and inclusion of ready-to-use kits with labeled standards and analysis software make it accessible for both research and clinical applications. Furthermore, the ability to monitor multiple viral proteins simultaneously and track infection dynamics or drug responses in real time positions this technology as a powerful tool for antiviral drug development, mechanistic studies, and clinical diagnostics, addressing a critical need for accurate and comprehensive herpesvirus monitoring.

Benefits

• High specificity and accuracy through direct quantification of viral proteins using signature peptides and targeted mass spectrometry.
• Multiplexed detection enabling simultaneous monitoring of 30–70% of the viral proteome across multiple herpesviruses in a single assay.
• Capability to track viral protein expression across all stages of infection (immediate-early, early, late), providing detailed temporal insights.
• Low sample input requirement (as few as 10,000 cells) facilitating use in limited or precious samples and enabling high-throughput scalability.
• Rapid and automated workflow with standardized data analysis and reporting, supporting reproducibility and ease of use across laboratories.
• Platform flexibility compatible with various mass spectrometry instruments, enhancing accessibility and adaptability.
• Applications in drug screening, mechanistic studies, clinical diagnostics, and research, supporting antiviral development and infection monitoring.
• Availability of ready-to-use kits and service models to streamline implementation and commercialization.

Applications

• High-throughput antiviral drug screening
• Clinical herpesvirus infection diagnostics
• Mechanistic studies of viral replication
• Quantitative monitoring in research labs
• Customizable herpesvirus detection kits

Additional info

This platform employs parallel reaction monitoring (PRM) and LC-MS/MS to quantify signature peptides from herpesvirus proteins. Utilizing validated tryptic peptide libraries, the workflow involves protein digestion and mass spectrometry analysis. Quantification integrates multiple transitions, normalized to host controls and heavy isotope-labeled standards. This high-throughput system monitors up to 70% of the viral proteome across all replication stages, facilitating drug screening and mechanistic studies in samples as small as 10,000 cells.