Development of a New Tandem Ion Exchange and Size Exclusion Chromatography Method To Monitor Vaccine Particle Titer in Cell Culture Media
Open accessAbstract
A new tandem chromatography method was developed to directly measure the titers of various vaccine candidate molecules in cell culture without a prior purification step. The method utilized a strong anion exchange chromatography (IEC) column in tandem with a size exclusion chromatography (SEC) column to efficiently separate the nanoparticle and virus-like particle (VLP) vaccine molecules from host cell proteins and other components in the cell culture media. The dual (charge and hydrodynamic size) separation mode was deemed necessary to achieve good separation of the vaccine product for quantitation. The method development and quality assessment illustrated herein was focused on the influenza vaccine candidate H1ssF, a hemagglutinin (group 1) stabilized stem molecule fused to ferritin to form nanoparticles. This newly established method was then successfully applied to several vaccine candidate developmental projects, such as the hemagglutinin-ferritin (HAF) nanoparticle and encephalitic alphavirus VLP-based vaccines. This IEC-SEC strategy was established as a platform approach for direct titer measurement of novel vaccine molecules in cell culture.
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Ion-exchange chromatography is a promising purification method that purifies vaccine process products using physical methods without adding exogenous products.
“Previous research has confirmed that ion-exchange chromatography is a promising purification method. It purifies vaccine process products using physical methods without adding exogenous products.”
From Evaluation of the Efficacy of the Vaccine Production Process in Removing Residual Host Cell DNA from the Vero Cell Rabies Vaccine (2024) · cites this paper as [27]
The Abstract (page not provided) describes ion-exchange chromatography combined with size-exclusion chromatography separating vaccine particles from cell-culture components. But the method is for analytical titer measurement without prior purification, not a vaccine-product purification process. The Materials and Methods describe buffers and salt used in the assay; the paper does not establish the claim that purification occurs without adding exogenous products. (LLM, conf=0.88)
Experimental — accuracy not yet measuredShaddeau et al. used tandem ion-exchange and size-exclusion chromatography to separate vaccine components from host cell proteins and other components in cell media to detect vaccine titres.
“Shaddeau et al. used tandem ion-exchange and size-exclusion chromatography to separate vaccine components from host cell proteins and other components in cell media to detect vaccine titres [27].”
From Evaluation of the Efficacy of the Vaccine Production Process in Removing Residual Host Cell DNA from the Vero Cell Rabies Vaccine (2024) · cites this paper as [27]
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Research works printed but not linked: 18
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Cited by
Sorted by citations
Li, J., Pan, R., Yue, F., Gao, T., Wu, X., Shi, L., Wang, Y., Zhao, D., Lan, Z., Chen, H., et al. (2024) · Vaccines · doi:10.3390/vaccines12121379 · Cited by 8 · see citing works
- Authors
- Shaddeau, A. W., Schneck, N. A., Li, Y., Ivleva, V. B., Arnold, F. J., Cooper, J. W., & Lei, Q. P.
- Affiliations
- National Institutes of Health (US)
- Corresponding author
- Schneck, Nicole A.
- Lei, Q. Paula
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2019-04-29
- Volume
- 91
- Issue
- 10
- Pages
- 6430-6434
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- PMC
- PMC11040568
- OpenAlex
- W2942736834
- Cited by
- 12
- References
- 0
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 1 · 2023: 1 · 2022: 1 · 2021: 2 · 2020: 3 · 2019: 1
- FWCI
- 0.58
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Analytical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Division of Intramural Research, National Institute of Allergy and Infectious Diseases
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Physical Sciences › Chemistry › Spectroscopy › Analytical Chemistry and Chromatography
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Keywords
- tandem chromatography, anion exchange chromatography, size exclusion chromatography, cell culture media, virus-like particles, nanoparticle vaccine, influenza vaccination
- MeSH terms
- Animals, Cell Line, Chromatography, Gel, Chromatography, Ion Exchange, Culture Media, Mammals, Particle Size, Vaccines
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