Question

Use your understanding of direct and indirect ELISA to answer the following questions

Answer 1

Answer:

Indirect ELISA will be the best approach to test the gonadotropin according to example.

Explanation:

Now, the scenario is that we are working on ribonuclease A (RNase A) which is an antigen and against this we produced mouse monoclonal antibodies to RNase A (anti-RNase A).

Note: No other laboratories have these monoclonal antibodies.

So, it means we don't have commercially available antibodies against RNase A and we are producing antibodies.

According to this scenario we will use indirect ELISA because firstly, we use antigen (RNase A) and then monoclonal antibodies against RNase A and after that we will use anti-monoclonal RNase A antibodies (secondary antibodies) with substrate to produce color according to concentration of antibodies.

Note: Secondary antibodies (anti-monoclonal RNase A antibodies) can be produced against RNase A antibodies.

We can't use sandwich ELISA because there is not any lab who has monoclonal antibodies against RNase A. So, first we use antigen then its antibodies and then against these antibodies we use secondary antibodies to determine the effectiveness of monoclonal antibodies against RNase A.