Site pages
Current course
Participants
General
Topic 1
Topic 2
Topic 3
Topic 4
Topic 5
Topic 6
Topic 7
Topic 8
Topic 9
Topic 10
Topic 11
Topic 12
Topic 13
Topic 14
Topic 15
Topic 16
Topic 17
Topic 18
Topic 19
Topic 20
Topic 21
Topic 22
Topic 23
Topic 24
Topic 25
Topic 26
3.2.3. Components of ELISA
1. Antigen Any substance that stimulates an immune response is known as antigen. It is the target protein in the sample that binds to the antibody. 2. Antibody An antibody is a protein made in response to an antigen. Each antibody binds in the epitope region of its antigen. Two types of antibodies viz. monoclonal or polyclonal can be produced and used as the capture and detection antibodies. The monoclonal antibodies are used as detection antibody as they have monospecificity toward a single epitope and allows fine detection and quantitation of small differences in antigen. 3. Enzyme conjugate An enzyme conjugate is an antibody joined with an enzyme. The joining of the enzyme to antibody is called conjugation. The detection enzyme can be linked directly to the primary antibody or introduced through a secondary antibody that recognizes the primary antibody. The most commonly used enzyme labels are horseradish peroxidase (HRP) and alkaline phosphatase (AP). 4. Substrate A substrate is a compound that undergoes change. The substrate binds to active sites on the surface of enzyme and gets converted by the change of color. The choice of substrate depends upon the assay sensitivity and instrumentation available for signal-detection such as spectrophotometer, fluorometer or luminometer. The common ELISA substrates are:
|