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Dear Mabtech staff and researchers, I am conducting HBV Surface antigen (HBsAg)-specific B cell IgG ELISPOT assay using U-Cytech kit (CT780-PR5). Attached is the image of the result. This time PBMCs from some healthy vaccinated (i.e. positive for anti-HBs) donors and clinically uninfected donors with no history of vaccination (i.e. negative for serum HBsAg, anti-HBsAg and anti-HBcAg) were stimulated with rIL2 and R848 for five days. PVDF membrane plate (Millipore, MSIPS4510) was coated with four recombinant SHBs antigens, PBS or IgG capture antibodies. The problem is that some Ab-negative donors show clearly positive spots for HBsAg (the yellow circle). I would appreciate if you could give me any advice on the possible causes of this result and the way to improve this with your products. I also would like to ask if anybody has similar experiences. Thank you. Keisuke ELISPOT20210708.pdf