Detection of harmful epitopes in foods - Quantitative analysis: the Enzyme Linked Immune Sorbent Assay

7 important questions on Detection of harmful epitopes in foods - Quantitative analysis: the Enzyme Linked Immune Sorbent Assay

Which 3 different ELISA are there?

  • Direct competitive ELISA
  • Indirect competitive ELISA
  • Non-competitive sandwich ELISA

What is direct competitive ELISA?

Antigen competes with a known amount of enzyme-labelled antigen for binding to an antibody immobilised on a solid phase

What is the process of non-competitive ELISA?

  1. Antigen solution is incubated with the antibody coated solid phase, in the absence of competing antigens or antibodies
  2. The amount of antigen bound is detected using a second enzyme conjugates antibody (Ig-PO)
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How long does direct competitive ELISA take?

1-2 hours

What is an advantage of indirect competitive ELISA?

It does not require the present of conjugates antigen.

What is an advantage of non-competitive sandwich ELISA?

The test is more sensitive, but has the drawback that a positive reaction requires the ability of an antigen to bind two antibodies. This is generally not a problem with intact proteins, but fragments that may be generated during processing, can remain undetected.

What is a disadvantage and advantages of ELISA?

Disadvantage:
  • Not portable


Advantages:
  • Instrumentation requires is relatively simple
  • Analysis times can be optimised down to 20-30 min, by adjusting the concentrations of reactants used
  • The procedure can be completely automated

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