Abstract
Many therapeutic viruses, such as oncolytic viruses, vaccines, or gene therapy vectors, may be administered by the intravenous route to maximize their delivery to target tissues. Blood components, such as antibody, complement and blood cells (such as neutrophils, monocytes, T cells, B cells or platelets) may result in viral neutralization and therefore reduce the therapeutic efficacy. This protocol will describe an in vitro assay by which to test the interaction of viruses with blood components. The effect of various factors can be isolated through fractionation. While whole blood can offer the most physiologically relevant snapshot, plasma can investigate the effects of antibody in concert with complement, and heat inactivated plasma will interrogate the effect of antibody alone.
Keywords: Complement, Antibody, Neutralization, Virus
Materials and Reagents
Equipment
Procedure
Representative data
Figure 2. Infectious Vaccinia virus recovery after incubation with blood, plasma or heat inactivated plasma from one representative human donor. Data can be represented as either titer (Log10 pfu/ml) as in panel A or as a proportion of virus recovered from the input (PBS or DMEM) as in panel B.
Notes
Recipes
Acknowledgments
The protocol was originally published in Evgin et al. (2015). This work was supported by the Terry Fox Research Foundation, the Ontario Institute for Cancer Research, the Ottawa Regional Cancer Foundation and the Canadian Institute for Health Research.
References
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