RUAS
Department of Biotechnology (2021-22)
Immunology and Molecular Biology Laboratory
Experiment 5
SANDWICH ELISA
Introduction:
Enzyme linked immunosorbent assay or ELISA is a sensitive immunological technique to detect the
presence of a specific antigen (Ag) or antibody (Ab) in a biological %arnple. It utilizes the dual
properties of antibody molecules being specific in reactivity and their ability to be conjugated to
active molecules such as enzymes. An enzyme conjugated with an antibody reacts with a
chromogenic colourJess substrate to generate a coloured reaction product. ELISA is extensively used
for diagnostic purpose, which utilizes the dual properties. It requires an immobilized
antigen/antibody bound to a solid support (e.g. microtitre plate or membrane). There are different
types of ELISAs for the detection of a protein of interest in a given sample. One of the most common
ELISA is Sandwich ELISA which can measure the amount of antigen which is sandwiched between
two layers of antibodies i.e. capture and detection antibody.
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