B cell ELISPOT
The B cell ELISPOT assay has been developed to enumerate ex vivo antigen-specific B cells on basis of antibody production at the single cell level. The assay is useful in different fields of biomedical research including vaccine development, infection, drug treatment and autoimmune diseases. For example, the number of ASCs following an immunization can be used to predict the magnitude and longevity of protection against a specific infectious disease.
Activation of memory B cells
B cell ELISPOT method
Examples of the B cell ELISPOT assay
The picture on the left demonstrates the results of tetanus-specific IgG secreting B cells produced by 5x104 human PBMC/well in 7 hours. In this assay, silver staining reagents for spot detection and a transparent polystyrene-bottomed plate were used. To activate the memory B cells, cells were preincubated in vitro for 5 days in the presence of a polyclonal B cell stimulus.
The picture on the right is an example of total IgG1 secreting B cells produced by 1x105 mouse (Balb/c) splenocytes/well in 20 hours. Spots were visualized by enzymatic staining on a PVDF-membrane bottomed plate. Activation of memory B cells was accomplished by an in vitro preincubation period of 2 days in the presence of a polyclonal B cell stimulus.