Real-time monitoring of attenuated cytomegalovirus using Raman spectroscopy allows non-destructive characterization during flow

Affiliations

¹ Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN 47907, USA; Birck Nanotechnology Center, Purdue University, West Lafayette, IN 47907, USA; Bindley Bioscience Center, Purdue University, West Lafayette, IN 47907, USA.
² Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN 47907, USA; Birck Nanotechnology Center, Purdue University, West Lafayette, IN 47907, USA.
³ School of Mechanical Engineering, Purdue University, West Lafayette, IN 47907, USA.
⁴ Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN 47907, USA; Department of Computer Science, Purdue University, West Lafayette, IN 47907, USA.
⁵ Analytical Research and Development, Merck & Co., Inc., West Point, PA 19486, USA.
⁶ Birck Nanotechnology Center, Purdue University, West Lafayette, IN 47907, USA; School of Material Science Engineering, Purdue University, West Lafayette, IN 47907, USA.
⁷ Process Research and Development, MRL, Merck & Co., Inc., West Point, PA 19486, USA.
⁸ Bioprocess Drug Substance & Commercialization, Merck & Co., Inc., Kenilworth, NJ 07033, USA.
⁹ School of Mechanical Engineering, Purdue University, West Lafayette, IN 47907, USA. Electronic address: ardekani@purdue.edu.
¹⁰ Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN 47907, USA; Birck Nanotechnology Center, Purdue University, West Lafayette, IN 47907, USA; Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907, USA. Electronic address: msverma@purdue.edu.

PMID: 40782423
DOI: 10.1016/j.saa.2025.126761

Free article

Real-time monitoring of attenuated cytomegalovirus using Raman spectroscopy allows non-destructive characterization during flow

Shreya Milind Athalye et al. Spectrochim Acta A Mol Biomol Spectrosc. 2026.

Free article

. 2026 Jan 15:345:126761.

doi: 10.1016/j.saa.2025.126761. Epub 2025 Aug 7.

Affiliations

¹ Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN 47907, USA; Birck Nanotechnology Center, Purdue University, West Lafayette, IN 47907, USA; Bindley Bioscience Center, Purdue University, West Lafayette, IN 47907, USA.
² Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN 47907, USA; Birck Nanotechnology Center, Purdue University, West Lafayette, IN 47907, USA.
³ School of Mechanical Engineering, Purdue University, West Lafayette, IN 47907, USA.
⁴ Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN 47907, USA; Department of Computer Science, Purdue University, West Lafayette, IN 47907, USA.
⁵ Analytical Research and Development, Merck & Co., Inc., West Point, PA 19486, USA.
⁶ Birck Nanotechnology Center, Purdue University, West Lafayette, IN 47907, USA; School of Material Science Engineering, Purdue University, West Lafayette, IN 47907, USA.
⁷ Process Research and Development, MRL, Merck & Co., Inc., West Point, PA 19486, USA.
⁸ Bioprocess Drug Substance & Commercialization, Merck & Co., Inc., Kenilworth, NJ 07033, USA.
⁹ School of Mechanical Engineering, Purdue University, West Lafayette, IN 47907, USA. Electronic address: ardekani@purdue.edu.
¹⁰ Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN 47907, USA; Birck Nanotechnology Center, Purdue University, West Lafayette, IN 47907, USA; Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907, USA. Electronic address: msverma@purdue.edu.

PMID: 40782423
DOI: 10.1016/j.saa.2025.126761

Abstract

Real-time monitoring of viral particles can have a crucial impact on vaccine manufacturing and can alleviate public health challenges by supporting continuous supply. Spectroscopic methods such as Raman spectroscopy can provide rapid and non-invasive measurements. Here, we have developed a Raman spectroscopy-based tool to monitor the quality and quantity of viral particles in a continuous flow setup. We characterized the attenuated human cytomegalovirus (CMV) across a wide range of concentrations (1.45 × 10¹⁰ to 2.90 × 10¹¹ particles/mL) and flow rates (100 μm/s to 1000 μm/s) within a square quartz capillary. This process analytical technology (PAT) tool enables the detection of viral particles even at high flow rates such as 1000 μm/s. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and dynamic light scattering (DLS) demonstrated that the samples maintain their integrity even after laser exposure, reiterating the non-invasive nature of Raman spectroscopy. To the best of our knowledge, this is the first report on characterizing CMV particles using Raman spectroscopy, especially under flow conditions. We have also demonstrated the limit of detection (LOD_min) (2.01 × 10¹⁰ particles/mL) for CMV particles in continuous flow (1000 μm/s) (via the Raman spectroscopy method), addressing the effects of flow rate, concentration, and sample integrity. This technology could enable process control in the bio-manufacturing of vaccines.

Keywords: Continuous manufacturing; Cytomegalovirus; In-situ analysis; Process analytical technology; Raman spectroscopy.

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Conflict of interest statement

Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Mohit S. Verma and Arezoo M. Ardekani report financial support was provided by National Institute for Innovation in Manufacturing Biopharmaceuticals (NIIMBL). Mohit S. Verma and Arezoo M. Ardekani report financial support was provided by U.S. Department of Commerce, National Institute of Standards and Technology. Mohit S. Verma and Arezoo M. Ardekani reports financial support was provided by Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc. Mohit S. Verma reports a relationship with Krishi, Inc. that includes: board membership, equity or stocks, and funding grants. Shreya M. Athalye, Murali K. Maruthamuthu, Ehsan Esmaili, Miad Boodaghidizaji, Arezoo Ardekani, and Mohit S. Verma have patent pending to Purdue Research Foundation. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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Real-time monitoring of attenuated cytomegalovirus using Raman spectroscopy allows non-destructive characterization during flow

Affiliations

Real-time monitoring of attenuated cytomegalovirus using Raman spectroscopy allows non-destructive characterization during flow

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Conflict of interest statement

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