Flow Virometry Sorting, B cell Cloning and Evaluation - CytoFLEX SRT and Viral Research

Virus-like particles (VLPs) are nanostructures that possess diverse applications in therapeutics, immunization, and diagnostics. They consist of one or more different molecules, have the ability to self-assemble, and are virus particle mimics in that they share the same form and size. However, they lack the proper genetic material to be infectious. Recently, VLPs have garnered increased attention in relation to SARS-CoV-2 vaccination. Due to their ability to elicit an immune response, VLPs have become an attractive alternative to conventional vaccination methods. However, contaminants from host cell production are a major issue. Using a combination of flow virometry, as described by Tang et al, and nano-flow cytometry, one can isolate and characterize VLPs more efficiently using the CytoFLEX SRT benchtop cell sorter to selectively capture VLPs of interest. In addition, upon vaccination, it has become tantamount to measure immune response and the associated pathways. Within the mechanism of SARS-CoV-2 vaccines, both follicular helper T (Tfh) and follicular regulatory T (Tfr) cells have an effect on germinal center (GC) reactions. GC reactions generate antibodies and memory B cells, and are regulated by a balance of Tfh and Tfr response. In order to study this delicate response, single B cell clones from SARS-CoV-2 immunized mice were sorted into 96 well plates using CytoFLEX SRT benchtop cell sorter prior to further downstream applications. Specificity, affinity, and somatic hypermutation were assessed via BCR sequencing and ELISA.

 

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