You decide to design an assay for flu infection detection. ha protein is only present on the surface of influenza viral particles so you choose to design an elisa assay to detect the presence of ha in human blood samples.

you will use three different antibodies for the assay. the experimental procedure is as follows:

step 1. incubate your assay plate with antibody (i) solution overnight to coat the surface of the assay plate with antibody (i). remove antibody (i) solution and wash the plate extensively.

step 2. add blood samples to the coated plate and incubate for 30 min. remove blood sample and wash the plate extensively.

step 3. add antibody (ii) solution to the plate and incubate for 30 min. remove antibody (ii) solution and wash the plate extensively.

step 4. add antibody (iii) solution to the plate and incubate for 30 min. remove antibody (iii) solution and wash the plate extensively.

step 5. add tmb substrate to the plate and monitor color development.