The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity

The overall goal of this ELISpot methodology is to allow the quantification of Human Antibody-secreting B cells from the blood. This method can help answer key questions in the bio-medical field about the immunology of B cells and vaccine research. The main advantage of this technique is that it allows the detection and measurement of anti-body secreting B cells at the single cell level.

Demonstrating the procedure will be Tsung-Chih Tseng, a graduate student from my laboratory. Immediately after obtaining a 10 ml blood sample from median cubital vein into a 15 ml tube containing potassium supplemented EDTA, invert the tube several times to prevent clot formation. Then add 35 ml of autoclaved red blood cell lysis buffer to the cells for a five minute incubation at room temperature.