Quantification of viral particles in adenovirus vector-based vaccines by nano-flow cytometry.
- Author:
Zhuowei SHI
1
;
Ying ZHANG
1
;
Qingya TIAN
1
;
Ziqiang WANG
1
;
Hong SHAO
1
Author Information
- Publication Type:Journal Article
- Keywords: OD260 method; adenovirus vector-based vaccine; nano-flow cytometry; particle size; viral particles
- MeSH: Adenoviridae/genetics*; Genetic Vectors; Flow Cytometry/methods*; Virion/isolation & purification*; Particle Size; Nanoparticles; Viral Vaccines
- From: Chinese Journal of Biotechnology 2025;41(8):3155-3164
- CountryChina
- Language:Chinese
- Abstract: This study aims to establish a method for counting the viral particles in adenovirus vector-based vaccines. Nano-flow cytometry was employed to analyze the viral particles in adenovirus-based vector vaccines at the single-particle level. Monodisperse silica nanoparticles with a refractive index close to that of the virus were selected as the particle size standard to calculate the viral particle size, which was then compared with the results obtained from transmission electron microscopy (TEM) to determine the gating strategy. Subsequently, a particle count standard was employed to calculate the viral particle concentration. The established method demonstrated good linearity, accuracy, precision, and specificity. The results of determined viral particle concentration showed a good correlation with the infectious titer. Compared with the conventional OD260 method, nano-flow cytometry can directly measure the viral particle concentration and indicate whether the sample has been disassembled according to changes in viral particle concentration and size, thus more accurately reflecting the actual infectious potency of the sample. The novel quantification method established in this study is capable of indicating the efficacy of adenovirus vector-based vaccines and provides effective technical support for the quality control of such products.
