Standardization and validation of Vero cell assay for potency estimation of diphtheria antitoxin serum.
Abstract: Diphtheria toxin has the capacity to block protein synthesis in cultured mammalian cells, and thus causing cell death. This capacity of diphtheria toxin was utilized for in-vitro neutralization test to determine antibody titer, using Vero cells, which have been found to be susceptible to diphtheria toxin. In the present study, a Vero cell assay was standardized and validated for potency estimation of diphtheria antitoxin serum (DATS). The results obtained by Vero cell assay were compared with in-vivo biological assay. High degree of correlation (+0.98) was found between in-vivo biological assay and in-vitro Vero cell assay. The assay has also been found to be effective in determining the rising antibody titer in the equines inducted in DATS production. The present study indicated that although biological assays hold the key for final potency estimations till date but in the future scenario in-vitro Vero cell assay may be a good alternative to in-vivo biological assay.
Publication Date: 2009-06-18 PubMed ID: 19540135DOI: 10.1016/j.biologicals.2009.05.002Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Validation Study
Summary
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This research was aimed at testing and confirming the effectiveness of the Vero cell assay for determining the potency of diphtheria antitoxin serum (DATS). Results were compared with the in-vivo biological assay, and a high correlation was found, indicating that the Vero cell assay could be a more suitable alternative in future.
Testing the Diphtheria Antitoxin Serum
- The study was built on the deployment of diphtheria toxin’s capacity to prevent protein synthesis in mammalian cells, causing cell death. This lethal effect of the toxin was used in an in-vitro experiment to discern the antibody titer using Vero cells, which have proven to be vulnerable to diphtheria toxin.
- The focus of the study was the standardization and validation of a Vero cell assay, to estimate the potency of the diphtheria antitoxin serum (DATS).
Comparing Vero Cell Assay with In-Vivo Biological Assay
- The experiment compared the results produced by the Vero cell assay with those produced by the in-vivo biological assay. This comparison was essential to ascertain if the Vero cell assay could deliver reliable results that would match or even surpass the in-vivo biological assay’s reliability.
- A high degree of correlation, represented by +0.98, was recorded between the in-vivo biological assay and the Vero cell assay. This relationship significantly indicates that the initial serum potency could either predict or have a significant bearing over the results of the latter, or vice versa.
The Effectiveness of the Vero Cell Assay
- The research verifies that the Vero cell assay can effectively determine a rise in the antibody titer within equines that were induced in the production of DATS. This effectiveness empowers the assay as an effective tool for serological testing or immunogenic potency estimation.
- The research concludes that although biological assays are still paramount for final potency estimation at present, the Vero cell assay could be considered a credible alternative in the future based on the high degree of correlation with the in-vivo biological assay.
Cite This Article
APA
Kumar S, Kanwar S, Bansal V, Sehgal R.
(2009).
Standardization and validation of Vero cell assay for potency estimation of diphtheria antitoxin serum.
Biologicals, 37(5), 297-305.
https://doi.org/10.1016/j.biologicals.2009.05.002 Publication
Researcher Affiliations
- Central Research Institute, Kasauli, Himachal Pradesh 173204, India. sunilk63@rediffmail.com
MeSH Terms
- Animals
- Antibody Specificity
- Chlorocebus aethiops
- Diphtheria Antitoxin / analysis
- Diphtheria Antitoxin / immunology
- Diphtheria Antitoxin / therapeutic use
- Female
- Guinea Pigs
- Horses
- Immune Sera / analysis
- Immune Sera / immunology
- Male
- Models, Biological
- Neutralization Tests / methods
- Neutralization Tests / standards
- Reference Standards
- Sensitivity and Specificity
- Temperature
- Time Factors
- Vero Cells
Citations
This article has been cited 1 times.- Khan MA, Singh R, Siddiqui S, Ahmad I, Ahmad R, Upadhyay S, Barkat MA, Ali AMA, Zia Q, Srivastava A, Trivedi A, Husain I, Srivastava AN, Mishra DP. Anticancer potential of Phoenix dactylifera L. seed extract in human cancer cells and pro-apoptotic effects mediated through caspase-3 dependent pathway in human breast cancer MDA-MB-231 cells: an in vitro and in silico investigation.. BMC Complement Med Ther 2022 Mar 15;22(1):68.
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