Basic Principles of ELISA

Abstract

This chapter introduces the basic test formats for the performance of solid-phase heterogeneous ELISA. This form of ELISA has been so useful because:

• 1. Substances, e.g., antibodies or antigens, may be passively adsorbed to solid surfaces, such as plastics. Microtiter plates in a 96-well format are commercially available for use in ELISA, along with suitable equipment for easy manipulation and dispensing of reagents. This allows use of small volumes and gives the ELISA the potential of handling high numbers of samples rapidly.

• 2. Since one of the reactants in the ELISA is attached to a solid-phase, the separation of bound and free reagents is easily made by simple washing procedures.

• 3. The result of an ELISA is a color reaction that can be observed by eye and read rapidly using specially designed multichannel spectrophotometers. This allows data to be stored and analyzed statistically.