How could you modify the Jerne Plaque assay in order to be a

How could you modify the Jerne Plaque assay in order to be able to enumerate single antibody producing cells in the above population of unsorted spleen cells?

Solution

In Jerne plaque assay, sheep\'s RBCs are taken and added to spleen cells. Antibodies will be produced. Then spleen cells suspension is plated onto molten agar via pour plate method. The agar plates are kept in incubator at 37degree C for one hour. The antibody producing cells will release antibody, and this antibody will react with antigen, i.e. the sheep\'s RBC. Then complement is added. RBCs will be lysed resulting in production of plague which can be counted.

Modification (to find out single antibody producing cells):

0.1 ml of sheep\'s RBC and 0.1ml of chicken RBC was added to the spleen cell suspension with molten agar. If antibody is produced against both sheep\'s and chicken\'s RBC, then a zone of complete lysis would occur around this cell.

If the cell produced antibody to only one species of RBC (i.e., one antigen-please note) the plaques would appear hazy because of the presence of the other type of antigen i.e. chicken RBC. These cells are single antibody producing cells.