Enzyme-Linked Immunosorbent Assay (ELISA)

Direct ELISA

Antigen is immobilized on walls of each well. Plate is then washed with PBS to
remove excess unbound antigens.

A blocking solution is added to prevent non-specific binding of the antibodies to the

walls of the well. Wash with PBS once again.

An antibody conjugated to an enzyme such as HRP is then added to each well,

depending on which component of the virus is being analyzed.

Add the detection substrate to each well; if the corresponding antigen is present in
the well, then the conjugated antibody will be bound to it, and the enzyme will
convert the substrate into a coloured product.

The more the antigens present, the more the conjugated antibodies that will bind
and the greater the enzyme activity, producing a darker colour.

Tips

- Teach groups how to use a pipette.

- Setup correct work-flow for stations.
- Label wells and pipettes to avoid getting confused.
- Change pipette tips for every well to avoid cross-contamination.