Version 1
: Received: 13 March 2024 / Approved: 13 March 2024 / Online: 14 March 2024 (15:33:09 CET)
How to cite:
Hotter, D.; Kunzelmann, M.; Kiefer, F.; Leukhardt, C.; Fackler, C.; Jaeger, S.; Solzin, J. High Throughput Determination of Infectious Virus Titers by Kinetic Measurement of infection-Induced Changes in Cell Morphology. Preprints2024, 2024030809. https://doi.org/10.20944/preprints202403.0809.v1
Hotter, D.; Kunzelmann, M.; Kiefer, F.; Leukhardt, C.; Fackler, C.; Jaeger, S.; Solzin, J. High Throughput Determination of Infectious Virus Titers by Kinetic Measurement of infection-Induced Changes in Cell Morphology. Preprints 2024, 2024030809. https://doi.org/10.20944/preprints202403.0809.v1
Hotter, D.; Kunzelmann, M.; Kiefer, F.; Leukhardt, C.; Fackler, C.; Jaeger, S.; Solzin, J. High Throughput Determination of Infectious Virus Titers by Kinetic Measurement of infection-Induced Changes in Cell Morphology. Preprints2024, 2024030809. https://doi.org/10.20944/preprints202403.0809.v1
APA Style
Hotter, D., Kunzelmann, M., Kiefer, F., Leukhardt, C., Fackler, C., Jaeger, S., & Solzin, J. (2024). High Throughput Determination of Infectious Virus Titers by Kinetic Measurement of infection-Induced Changes in Cell Morphology. Preprints. https://doi.org/10.20944/preprints202403.0809.v1
Chicago/Turabian Style
Hotter, D., Stefan Jaeger and Johannes Solzin. 2024 "High Throughput Determination of Infectious Virus Titers by Kinetic Measurement of infection-Induced Changes in Cell Morphology" Preprints. https://doi.org/10.20944/preprints202403.0809.v1
Abstract
Infectivity assays are the key analytical technology for development of manufacturing processes for virus-based therapeutics. Here, we introduce a novel assay format that utilizes label-free bright field images to determine the kinetics of infection-dependent changes in cell morphology. In particular, cell rounding is directly proportional to the amount of infectious virus applied, enabling rapid determination of viral titers in relation to a standard curve. Our kinetic infectious virus titer (KIT) assay is stability-indicating and, due to its sensitive readout method, provides results within 24 hours post-infection. Compared to traditional infectivity assays, which depend on a single readout of an infection endpoint, cumulated analysis of kinetic data by a fit model results in precise results (CV < 20%) based on only three wells per sample. This approach allows for a high throughput with ~400 samples processed by a single operator per week. We demonstrate the applicability of the KIT assay for VSV-GP and NDV, but it can potentially be extended to a wide range of viruses that induce morphological changes upon infection. The versatility of this assay, combined with its independence from specific instruments or software, makes it a promising solution to overcome the analytical bottleneck in infectivity assays within the pharmaceutical industry and as a routine method in academic research.
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
