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Pharmacology-II Notes(The Cellular level of organisation)Disadvantages of competitive ELISA ELISA advantages and disadvantages ppt

Advantages and Disadvantages of each ELISA type

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Advantages Disadvantages
Direct ELISA
  1. Simple protocol, time-saving, and reagents-saving.
  2. No cross-reactivity from secondary antibody.
  1. High background.
  2. No signal amplification, since only a primary antibody is used and a secondary antibody is not needed.
  3. Low flexibility, since the primary antibody must be labeled.
Indirect ELISA
  1. Signal amplification, since one or more secondary antibodies can be used to bind to the primary antibody.
  2. High flexibility, since the same secondary antibody can be used for various primary antibodies.
  1. Complex protocol compared with direct ELISA.
  2. Cross-reactivity from secondary antibody.
Sandwich ELISA
  1. High flexibility.
  2. High sensitivity.
  3. High specificity, since different antibodies bind to the same antigen for detection.
  1. The antigen of interest must be large enough so that two different antibodies can bind to it at different epitopes.
  2. It’s sometimes difficult to find two different antibodies that recognize different epitopes on the antigen of interest and cooperate well in a sandwich format.
Competitive ELISA
  1. High flexibility.
  2. High sensitivity.
  3. Best for the detection of small antigens, even when they are present in low concentrations.
  1. Relatively complex protocol.
  2. Needs the use of inhibitor antigen.
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