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Determination of lentiviral titer by surface enhanced Raman scattering
Courtney J Morder1, Brian T Scarpitti1, Karin M Balss2
1Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA. schultz.133@osu.edu.
Analytical Methods : Advancing Methods and Applications
|March 11, 2022
Summary
Surface-enhanced Raman spectroscopy (SERS) offers a rapid method for determining lentiviral titer, crucial for gene therapy and vaccine development. This technique accurately quanties virus particles, overcoming limitations of traditional cell culture methods.
Area of Science:
- Biotechnology
- Spectroscopy
- Virology
Background:
- Lentiviruses are essential tools in biomedical applications like gene therapy and vaccine development.
- Accurate quantification of lentiviral infectious titer is critical for these applications.
- Current methods relying on cell culture are time-consuming and labor-intensive.
Purpose of the Study:
- To explore surface-enhanced Raman spectroscopy (SERS) as a rapid method for determining lentiviral titer.
- To assess the sensitivity of SERS to genetic modifications within lentiviral vectors.
- To develop a SERS-based model for accurate viral titer quantification.
Main Methods:
- Lentiviruses with and without a GFP gene were analyzed using SERS.
- Plasmonic metallic nanostructures were employed to amplify Raman signals.
- Chemometric analysis, specifically multivariate curve resolution (MCR), was used to correlate SERS spectra with viral concentration.
- SERS performance was evaluated using different metals and excitation wavelengths.
Main Results:
- SERS demonstrated sensitivity to the presence of the GFP gene in lentiviral vectors.
- MCR analysis successfully identified a spectral component correlating with GFP-containing lentivirus concentration.
- The viral titer determined by SERS showed strong agreement with results from traditional GFP expression analysis in infected cells.
Conclusions:
- SERS provides a rapid and accurate alternative to traditional methods for determining lentiviral titer.
- This technique is sensitive to specific genetic components within lentiviruses.
- SERS holds significant promise for accelerating research and development in lentiviral vector applications.
