Flow cytometric monitoring of influenza A virus infection in MDCK cells during vaccine production

Abstract

Background: In cell culture-based influenza vaccine production the monitoring of virus titres and cell physiology during infection is of great importance for process characterisation and optimisation. While conventional virus quantification methods give only virus titres in the culture broth, data obtained by fluorescence labelling of intracellular virus proteins provide additional information on infection dynamics. Flow cytometry represents a valuable tool to investigate the influences of cultivation conditions and process variations on virus replication and virus yields.

Modeling the Intracellular Dynamics of Influenza Virus Replication To Understand the Control of Viral RNA Synthesis

“…Experimentally, protein synthesis was found to proceed unhindered even at late times postinfection (26), and for M1, the most abundant component in a virus particle, accumulation in cells was shown even 24 hpi despite significant virus release (66). Similarly, strong M1 and NP fluorescence could be detected by flow cytometry at late times postinfection (52). Hence, it seems that the abundance of viral components is sufficient to release more virus particles, and that other processes constitute a bottleneck.…”

Population balance modeling of influenza virus replication in MDCK cells during vaccine production

“…With the help of flow cytometry it is possible to systematically collect data of various properties that characterize heterogeneous cell populations [Nichols et al, 1993;Srienc, 1999;Schulze-Horsel et al, 2008]. In Fig.…”

“…In Fig. 9 initial cell distributions are shown together with the respective experimental data recorded by Schulze-Horsel et al [2008] for the infection of adherent MDCK cells with equine influenza A/Newmarket/1/93 (H3N8).…”

“…However, uninfected cells and infected cells which are currently in the latent phase are considered to keep their initial fluorescence intensity. In the measurements performed by Schulze-Horsel et al [2008] the specific contributions of the two proteins M1 and NP to the overall fluorescence intensity are not discriminated. Hence, in the model both proteins are not discriminated either and their production is merged to a general production of viral protein.…”

A microfluidic platform for real-time and in situ monitoring of virus infection process

“…1,2 It is fundamental to understand the virus infection process for effectively reducing the damage caused by viruses. For most virus detection techniques, such as polymerase chain reaction (PCR), western blot, flow cytometry, and transmission electron microscopy (TEM), [3][4][5][6][7] the virus samples are often obtained from the infection system at a certain time point, which interrupts the virus infection process. These techniques cannot be applied to monitor the dynamic behaviors of virus infection in real time and in situ.…”