Skip to main content

Viral Infectivity Quantification and Neutralization Assays Using Laser Force Cytology

  • Protocol
  • First Online:
Vaccine Delivery Technology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2183))

Abstract

Despite the widespread need to assess cell-based viral infectivity during vaccine development and production, as well as viral clearance monitoring and adventitious agent testing for viral safety, traditional methods, including the end-point dilution assay (TCID50) and viral plaque assay, are slow, labor-intensive, and can vary depending upon the skill and experience of the user. LumaCyte’s Radiance® instrument uses Laser Force CytologyTM (LFC), a combination of advanced optics and microfluidics, to rapidly analyze the viral infectivity of cell populations in a quantitative fashion. LFC applies optical and fluidic forces to single cells in order to measure their intrinsic biophysical and biochemical properties without the use of stains, antibodies or fluorescent labels. These properties, including refractive index, change with a wide variety of biological phenomena, including viral infection, cell differentiation, activation, size, and cytoskeletal stiffness. Here, we present the experimental design and methods to use LFC data to facilitate rapid and robust infectivity measurements for a variety of applications including initial titer measurement (TCID50 replacement), in-process infectivity (e.g., bioreactor monitoring), and viral neutralization (PRNT replacement).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. World Health Organization (1997) Manual of laboratory methods for testing of vaccines used in the WHO Expanded Programme on Immunization

    Google Scholar 

  2. Guidance for Industry (2010) Characterization and qualification of cell substrates and other biological materials used in the production of viral vaccines for infectious disease indications. U.S.D.o.H.a.H. Services, editor. Office of Communication, Outreach and Development (OCOD) (HFM-40)

    Google Scholar 

  3. Gombold J et al (2014) Systematic evaluation of in vitro and in vivo adventitious virus assays for the detection of viral contamination of cell banks and biological products. Vaccine 32(24):2916–2926

    Article  Google Scholar 

  4. Gates IV et al (2009) Quantitative measurement of varicella-zoster virus infection by semiautomated flow cytometry. Appl Environ Microbiol 75(7):2027–2036

    Article  CAS  Google Scholar 

  5. Donis RO et al (2014) Performance characteristics of qualified cell lines for isolation and propagation of influenza viruses for vaccine manufacturing. Vaccine 32(48):6583–6590

    Article  CAS  Google Scholar 

  6. Shurtleff AC et al (2012) Standardization of the filovirus plaque assay for use in preclinical studies. Viruses 4(12):3511–3530

    Article  Google Scholar 

  7. Chou SW, Scott KM (1988) Rapid quantitation of cytomegalovirus and assay of neutralizing antibody by using monoclonal antibody to the major immediate-early viral protein. J Clin Microbiol 26(3):504–507

    Article  CAS  Google Scholar 

  8. Thomas JA et al (2016) Development and application of a simple plaque assay for the human malaria parasite Plasmodium falciparum. PLoS One 11(6):e0157873

    Article  Google Scholar 

  9. Heppner DG Jr et al (2017) Safety and immunogenicity of the rVSVG-ZEBOV-GP Ebola virus vaccine candidate in healthy adults: a phase 1b randomised, multicentre, double-blind, placebo-controlled, dose-response study. Lancet Infect Dis 17(8):854–866

    Article  CAS  Google Scholar 

  10. Salje H et al (2014) Variability in dengue titer estimates from plaque reduction neutralization tests poses a challenge to epidemiological studies and vaccine development. PLoS Negl Trop Dis 8(6):e2952

    Article  Google Scholar 

  11. Hebert CG et al (2018) Rapid quantification of vesicular stomatitis virus in Vero cells using Laser Force Cytology. Vaccine 36(41):6061–6069

    Article  Google Scholar 

  12. Hebert CG, Hart SJ, Terray A (2014) Label free detection of pseudorabies virus infection in Vero cells using laser force analysis. Analyst 139(6):1472–1481

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Colin G. Hebert .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Hebert, C.G., Rodrigues, K.L., DiNardo, N., Hachmann, AB. (2021). Viral Infectivity Quantification and Neutralization Assays Using Laser Force Cytology. In: Pfeifer, B.A., Hill, A. (eds) Vaccine Delivery Technology. Methods in Molecular Biology, vol 2183. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0795-4_34

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0795-4_34

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0794-7

  • Online ISBN: 978-1-0716-0795-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics