ORIGINAL RESEARCH article
Front. Bioeng. Biotechnol.
Sec. Bioprocess Engineering
Volume 13 - 2025 | doi: 10.3389/fbioe.2025.1611428
Development and validation of a qPCR assay for detection of residual host cell DNA for rabies vaccines produced in Vero cells
Provisionally accepted
- 1National Institutes for Food and Drug Control (China), Beijing, Beijing Municipality, China
- 2Huzhou Shenke Biotechnology Co., Ltd, Huzhou, China
Select one of your emails
You have multiple emails registered with Frontiers:
Notify me on publication
Please enter your email address:
If you already have an account, please login
You don't have a Frontiers account ? You can register here
Residual host cell DNA in biological products poses potential risks of tumorigenicity and infectivity. Therefore, current regulatory authorities including the World Health Organization (WHO), and the U.S. Food and Drug Administration (FDA) have limited the acceptance amounts of residual DNA (not more than 10 ng per dose). Among various detection methods for residual DNA, quantitative PCR (qPCR) is recognized as the most practical method for its high sensitivity, accuracy, precision, and efficiency. This study aimed to develop a qPCR method for detecting residual Vero DNA to facilitate downstream purification and ensure quality control during vaccine manufacturing. We selected two highly repetitive Vero genomic DNA sequences as amplification targets: the "172 bp" sequence and the Alu repetitive sequence. The qPCR method was meticulously optimized and validated for linearity, quantitation limit, detection limit, specificity, accuracy, repeatability, intermediate precision, and robustness. The "172 bp" sequence qPCR assay showed excellent linearity and amplification efficiency, establishing a dependable approach for quantifying residual Vero DNA in pharmaceuticals and for regulatory compliance monitoring.
Keywords: Vero cell, rabies vaccine, Exogenous DNA residue, qPCR, Method validation
Received: 14 Apr 2025; Accepted: 29 Aug 2025.
Copyright: © 2025 Zhao, Zong, Wu, Yuhua, Wu, Yang and Cao. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Zhixing Yang, Huzhou Shenke Biotechnology Co., Ltd, Huzhou, China
Shouchun Cao, National Institutes for Food and Drug Control (China), Beijing, Beijing Municipality, China
Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.