Development of a Vero cell DNA reference standard for residual DNA measurement in China
Open accessAbstract
This collaborative study developed a Vero cell DNA reference for standardizing dot blot hybridization, an assay widely employed to measure residual DNA contents of viral vaccines prepared with Vero cells. High purity of Vero cell DNA was extracted and characterized by Hind III enzyme digestion and DNA sequencing. Then, with a cooperative calibration, the concentration of Vero cell DNA reference bulk solution was determined (64.0 ± 1.9 μg/mL, OD 260/OD 280 = 1.87) and diluted (40 ng/mL) with Tris-EDTA buffer containing bovine serum albumin as freeze-dried excipients. With industrial filling apparatus, the diluted bulk was loaded into ampoules (0.5 mL each) which were heat sealed after nitrogen filling. Finally, a collaborative study showed that the Vero cell DNA reference could reach a sensitivity of 1 to 5 pg/dot and maintained good stability after accelerated destruction test. The successful establishment of the Vero cell DNA quantitative reference will facilitate the standardization of dot blot hybridization for testing residual host cell DNA.
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The AGMr(HindIII)-1 fragment is the unique sequence of the Vero cell genome, and the fragment distribution of this sequence and its derivatives is broad-spectrum representative of Vero cells and can be used for quantitative analysis of HCD.
“This fragment is the unique sequence of the Vero cell genome, and the fragment distribution of this sequence and its derivatives is broad-spectrum representative of Vero cells and can be used for the quantitative analysis of HCD [38,39].”
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 [39]
Cites
Singer, D. S. (1979) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)50624-4 · Cited by 57 · cited 1× · see citing works
“This pattern of cleavage was consistent with previous report.(5) Finally, the origin of DNA was validated by DNA sequencing.”
ResultKline, M. C., Duewer, D. L., Travis, J. C., Smith, M. V., Redman, J. W., Vallone, P. M., Decker, A. E., & Butler, J. M. (2009) · Analytical and Bioanalytical Chemistry · doi:10.1007/s00216-009-2782-0 · Cited by 44 · see citing works
18 – Rabies Vaccine Produced in Cell Culture: Production Control and Clinical Results
Roumiantzeff, M., Ajjan, N., Branche, R., Fournier, P., Montagnon, B., Trotemann, P., & Vincent-Falquet, J. (1984) · doi:10.1016/b978-0-12-429601-5.50023-x · Cited by 16
Niimi, S., Oshizawa, T., Naotsuka, M., Ohba, S., Yokozawa, A., Murata, T., & Hayakawa, T. (2002) · Biologicals (Print) · doi:10.1006/biol.2001.0318 · Cited by 13
Gao, K., Rao, C.-M., Tao, L., Han, C.-M., Shi, X.-C., Wang, L., Fan, W.-H., Yu, L., & Wang, J.-Z. (2012) · Biologicals (Print) · doi:10.1016/j.biologicals.2011.08.013 · Cited by 5
Self-citationA collaborative study on DNA quantitation in biological products.
Robertson, J., & Heath, A. (1995) · Biologicals (Print) · doi:10.1006/biol.1995.0033 · Cited by 4 · cited 1×
“The variability can be attributed to numerous factors that are introduced in the multiple steps of the experimental procedure.(4) An important but often ignored factor, however, is the absence of a Vero cell DNA reference standard.”
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Progress on the research and development of inactivated EV71 whole-virus vaccines
Liang, Z.-L., Mao, Q.-Y., Wang, Y., Zhu, F., Li, J., Yao, X., Gao, F., Wu, X., Xu, M., & Wang, J.-Z. (2013) · Human Vaccines & Immunotherapeutics · doi:10.4161/hv.24949 · Cited by 57 · see citing works
Self-citationLi, 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
Self-citationWang, G.-J., & Shao, M. (2021) · Human Vaccines & Immunotherapeutics · doi:10.1080/21645515.2021.2002640 · Cited by 7 · see citing works
Development of a Reference Standard of Escherichia coli DNA for Residual DNA Determination in China
Wang, L., Rao, C.-M., Gao, K., Li, Y.-H., Fu, Z., Bi, H., & Wang, J.-Z. (2013) · PLoS ONE · doi:10.1371/journal.pone.0074166 · Cited by 7
“standard materials were referred to and strictly followed [3,18,19,20].”
BackgroundSelf-citationEffect of Fetal Bovine Serum Concentration towards Vero Cells Growth on Culture in DMEM Medium
Ikasari, B. N., Alfarizi, S., Fauziyah, S., Wardhani, P., Aryati, Soegijanto, S., & Sucipto, T. H. (2022) · Jurnal Teknologi Laboratorium · doi:10.29238/teknolabjournal.v11i2.313 · Cited by 4
“__________________ Conditions are created by regulating temperature, pH, oxygen, CO 2 , osmotic pressure, the surface to adhere to cells, nutrients, and vitamins, protection against toxic substances, hormones, and growth factors that regulate cell growth and differentiation.”
BackgroundZhao, D.-H., Zong, W., Wu, W., Li, Y.-H., Wu, Z., Yang, Z., & Cao, S. (2025) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2025.1611428 · Cited by 3 · see citing works
Self-citationShao, J., Gao, F., Lin, H., Mao, Q.-Y., Chen, P., Wu, X., Yao, X., Kong, W., & Liang, Z.-L. (2016) · PLoS ONE · doi:10.1371/journal.pone.0153867 · Cited by 3
“Inactivated EV71 whole-virus liquid bulk and cell lines Inactivated EV71 whole-virus liquid bulk (subgenotype C4, vero cell), containing 6000U/ml of Ag, less than 5EU/ml bacterial endotoxin, and with residual vero cell DNA below than 10pg per 400U antigen (Ag) measured by the dot blot hybridization assay following Cao…”
MethodRegulatory science accelerates the development of biotechnology drugs and vaccines by NIFDC
Liang, Z.-L., Mao, Q.-Y., Wang, Y., Li, C.-G., Gao, K., & Wang, J.-Z. (2014) · Emerging Microbes and Infections · doi:10.1038/emi.2014.66 · Cited by 3
Self-citationMărgineanu, E. (2021) · Circular Economy and Sustainability · doi:10.1007/s43615-021-00062-9 · Cited by 1
“1846001A in 2019) containing Vero cells, science literature acknowledge that this ingredient has a huge market share in viral vaccines which determines drug regulators to assess the residual DNA content of Vero cells in vaccines for human use, given the potential risk of carcinogenesis [7, 44].”
BackgroundZhao, D.-H., Wang, Y.-P., Wu, X.-H., Shi, L.-T., Li, J., Li, Y.-H., & Cao, S. (2026) · Frontiers in Microbiology · doi:10.3389/fmicb.2026.1708044
“…of the People’s Republic of China (2010 Edition, Part III), we successfully developed the national reference material for Vero cell DNA quantification applicable to the probe hybridization method (Cao et al., 2013), which was approved for use in 2012 (identification number: 250015–201101).”
Self-citation
- Authors
- Cao, S., Dong, G., Tang, J., Li, J., Liu, J., Shi, L., Li, C., & Wang, J.
- Affiliations
- National Institutes for Food and Drug Control (CN)
- Corresponding author
- Wang, Junzhi
- Venue
- Human Vaccines & Immunotherapeutics
- Publisher
- Informa UK Limited
- Published
- 2013-02
- Volume
- 9
- Issue
- 2
- Pages
- 413-419
- ISSN
- 2164-5515 (ISSN-L)
- 2164-554X
- PMC
- PMC3859766
- OpenAlex
- W2060409558
- Cited by
- 12
- References
- 18
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 1 · 2022: 1 · 2021: 2 · 2019: 1 · 2016: 1 · 2015: 1 · 2014: 1 · 2013: 2
- FWCI
- 0.75
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- cc-by-nc
- Locations
- Human Vaccines & Immunotherapeutics (publishedVersion, cc-by-nc)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Science and Technology Major Project
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Social Sciences › Decision Sciences › Statistics, Probability and Uncertainty › Scientific Measurement and Uncertainty Evaluation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
- Keywords
- dot blot, reference material development, collaborative study, stability testing, DNA quantification
- MeSH terms
- Animals, Chlorocebus aethiops, China, DNA, Nucleic Acid Hybridization, Quality Control, Reference Standards, Technology, Pharmaceutical, Vero Cells, Viral Vaccines
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