ELISA, standing for Enzyme-Linked ImmunoSorbent Assay, is a powerful and versatile technique employed in various immunological applications. It functions as a highly sensitive tool for detecting and quantifying the presence of specific molecules, typically antigens or antibodies, within a sample. The core principle of ELISA relies on the exquisite specificity of antigen-antibody interactions. Here's a breakdown of the key steps involved:

  1. Antigen Coating: The ELISA plate is coated with a capture molecule, which can be either the target antigen itself or an antibody specific to the target antigen. This initial step allows for the capture and immobilization of the target molecule on the plate's surface.
  2. Sample Introduction:  The sample containing the suspected antigen or antibody is then added to the ELISA plate.  If the target molecule is present in the sample, it will bind specifically to the captured molecule on the plate.
  3. Washing:  Unbound molecules are removed through a thorough washing step.
  4. Detection Antibody Addition:  An enzyme-conjugated "detector" antibody, specific for the target molecule or the previously bound antibody, is introduced. This detector antibody binds to the target molecule or the captured antibody-target molecule complex
  5. Substrate Addition and Signal Generation:  A substrate specific to the conjugated enzyme is then added.  The enzyme cleaves the substrate, generating a quantifiable signal, often colorimetric or luminescent. The intensity of this signal is directly proportional to the amount of target molecule present in the original sample.

Schematic presentation of basic types of ELISA

ELISA offers several advantages, including high sensitivity, specificity, and the ability to analyze multiple samples simultaneously.  Due to its versatility, ELISA finds applications in various fields, including:

  • Diagnosing infectious diseases: Detecting the presence of viral or bacterial antigens or antibodies in patient samples.
  • Allergy testing: Identifying specific allergens an individual might be sensitive to.
  • Autoimmune disease diagnosis: Detecting autoantibodies, which are antibodies that mistakenly target the body's own tissues.
  • Food safety testing: Identifying the presence of foodborne pathogens or allergens in food products.
  • Drug discovery: Monitoring drug levels in the body or screening for potential drug targets.

Overview of advantages and disadvantages of ELISA and LC-MS/MS for gluten detection.

Overall, ELISA remains a cornerstone technique in immunology, providing valuable insights into antigen-antibody interactions and aiding in disease diagnosis, research, and various other applications.

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