How do I increase the ELISA sensitivity?

The ELISA sensitivity is the lowest detection level of target antigen that can be measured using an ELISA kit. It is affected by a variety of factors, such as incubation temperature, incubation time, capture antibody concentration and immobilization, detection antibody specificity and secondary antibody usage. Sandwich ELISA offers higher sensitivity compared to direct or indirect ELISA.

In the sandwich ELISA, the primary step involves binding of antigen to the capture antibody. The next step in the ELISA is antibody detection. A monoclonal antibody is used to provide improved specificity. The addition of a biotinylated secondary antibody followed by conjugated streptavidin can be used to increase signal

Other parameters, such as incubation time and temperature, should be optimized empirically to achieve higher sensitivity.