sebbeols Posted January 31, 2017 Report Share Posted January 31, 2017 Hi Mabtech! I am working on optimising a B cell ELISpot for a vaccination study. I have previously worked with coating Ag directly on plates or utilising biotinylated protein Ag. For the current study there is unfortunately no protein Ag currently identified as an Ab/B cell target that we could use for the pathogens vaccinated for (different live attenuated vaccines). Thus, we only have the vaccine itself or formalin-fixed virions to work with. My attempts at coating directly with the vaccine have been fruitless and it may be because of the low concentration of virions in the vaccine (1.5-2.4 ug/0.5mL). In your brochure for the B cell ELISpot, you have a figure about the Pandemrix vaccine (coat vs biotin, Fig 4). Is the biotinylated Ag used there an HA protein or is it the actual vaccine that you have managed to biotinylate? Do you guys have any experience in biotinylating vaccines or virions for use as probes in ELISpot? Or do you believe that coating directly with the formalin-fixed virions is a better way to go (have not yet tried this)? Best regards, Sebastian Link to comment Share on other sites More sharing options...
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