The ELISA Test

• What does the acronym ELISA stand for?

  Enzyme-linked immunosorbent assay
• Why do we need to determine the quantity of a protein in biology?

  To assess levels of antigens in blood
• What are the stages of the ELISA test?

  1. Apply sample containing antigen to well
  2. Wash to remove unbound antigens
  3. Add specific antibody to bind antigen
  4. Wash to remove excess antibodies
  5. Add second antibody linked to enzyme
  6. Wash to remove unbound second antibody
  7. Add substrate for enzyme
  8. Measure color intensity to quantify antigen
• What is the purpose of washing the well after applying the sample?

  To remove unbound antigen molecules
• What is the role of the second antibody in the ELISA test?

  It binds to the first antibody and has an enzyme
• How does the enzyme in the second antibody contribute to the ELISA test?

  It converts substrate into a colored product
• What does the intensity of the color produced in the ELISA test indicate?

  It indicates the amount of antigen present
• Why do we use different dilutions of the sample in an ELISA test?

  To ensure all antigen molecules stick to the plate
• What problem does using a range of dilutions solve in the ELISA test?

  It prevents overloading of antigen in wells
• How are ELISA tests often automated in practice?

  By allowing many samples to be read accurately
• For what purpose is the ELISA test particularly valuable?

  Testing for infectious diseases like HIV
• What are the advantages of using ELISA tests in medical diagnostics?

  • Quantifies protein levels accurately
  • Can analyze multiple samples simultaneously
  • Automated processes increase efficiency
  • Useful for detecting infectious diseases
• What are monoclonal antibodies?

  Antibodies created to be identical
• What does "mono" mean in monoclonal antibodies?

  One
• What does "clonal" refer to in monoclonal antibodies?

  Identical
• How are monoclonal antibodies used in medical treatment?

  As drugs for targeted therapy
• What is the first type of monoclonal antibody therapy mentioned?

  Direct monoclonal antibody therapy
• How do direct monoclonal antibodies work in cancer treatment?

  They bind to cancer cell antigens
• What is the advantage of using monoclonal antibodies in cancer treatment?

  They target only cancer cells
• What is indirect monoclonal antibody therapy?

  Antibodies deliver cytotoxic drugs to cancer cells
• What are "bullet drugs" in the context of monoclonal antibodies?

  Drugs delivered directly to cancer cells
• What is one common use of monoclonal antibodies in medical diagnosis?

  Pregnancy tests
• What does the ELISA test stand for?

  Enzyme-linked immunosorbent assay
• How does a pregnancy test work?

  It detects a hormone in urine
• What happens when the mobile antibody binds to the hormone in a pregnancy test?

  It creates a visible line on the test
• What is the purpose of the control band in a pregnancy test?

  To confirm the test is working
• What is the first step in the ELISA test?

  Add the test sample to the device
• Why do you wash the device after adding the test sample in the ELISA test?

  To remove unbound test sample
• What is the role of the second antibody in the ELISA test?

  It binds to the first antibody
• What does the enzyme attached to the second antibody do in the ELISA test?

  Catalyzes a color change reaction
• What does a color change in the ELISA test indicate?

  The antigen is present in the sample
• How does the intensity of the color in the ELISA test relate to the antigen quantity?

  Darker color indicates more antigen present
• What ethical concern is associated with monoclonal antibody production?

  Use of animals in the process
• What happens to animals used in monoclonal antibody production?

  Their spleens are removed, leading to death
• Why is there an ethical debate regarding monoclonal antibodies?

  Is it justified to use animals for human benefit?
• What is one recent application of monoclonal antibodies?

  COVID-19 antibody tests
• What is the main function of antibodies?

  To bind specifically to antigens
• What is the significance of the binding site on antibodies?

  It is complementary to a specific antigen
• How do monoclonal antibodies differ from natural antibodies?

  They are produced in large identical quantities
• What is the role of the spleen in monoclonal antibody production?

  It produces antibodies in animals