Immunoelectrophoresis Overview
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Immunoelectrophoresis Overview

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Questions and Answers

What is the first step in preparing a sample for immunoelectrophoresis?

  • Diluting the sample with protein diluent solution (correct)
  • Adding the antiserum to the samples
  • Soaking the gel in saline solution
  • Applying the sample across the slits

    • How long does electrophoresis run during the immunoelectrophoresis process?

  • 20 minutes at 100 volts (correct)
  • 30 minutes at 150 volts
  • 15 minutes at 75 volts
  • 10 minutes at 50 volts

    • What does the presence of elliptical precipitin in immunoelectrophoresis indicate?

  • Successful antigen-antibody interaction (correct)
  • A faulty sample preparation
  • Improper electrophoresis conditions
  • No antigen-antibody interaction

    • What is an application of immunoelectrophoresis in clinical diagnostics?

    <p>Monitoring antigen and antibody purity</p> Signup and view all the answers

    What is the purpose of staining the gel during immunoelectrophoresis?

    <p>To enhance visibility of precipitin lines</p> Signup and view all the answers

    What characteristic is used to identify different proteins in immunoelectrophoresis?

    <p>Intensity, shape, and position of precipitation lines</p> Signup and view all the answers

    Which condition might immunoelectrophoresis help diagnose?

    <p>Hypogammaglobulinemia</p> Signup and view all the answers

    What is one of the advantages of using immunoelectrophoresis?

    <p>It can identify multiple antigens in serum</p> Signup and view all the answers

    What is one major limitation of immunoelectrophoresis compared to immunofixation electrophoresis?

    <p>It is harder to interpret.</p> Signup and view all the answers

    What does counter current immunoelectrophoresis require for the movement of antigen and antibody?

    <p>Electric field.</p> Signup and view all the answers

    What indicates a positive reaction in counter current immunoelectrophoresis?

    <p>Presence of a single precipitin line.</p> Signup and view all the answers

    What is a common application of counter current immunoelectrophoresis?

    <p>To detect antibodies for a specific antigen.</p> Signup and view all the answers

    In counter current immunoelectrophoresis, where do the antigen and antibody migrate?

    <p>Antigen towards the anode, antibody towards the cathode.</p> Signup and view all the answers

    What does the absence of a precipitin line in counter current immunoelectrophoresis indicate?

    <p>The absence of antigen-antibody interaction.</p> Signup and view all the answers

    What is a potential drawback of using immunoelectrophoresis in food analysis?

    <p>Limited availability of specific antibodies.</p> Signup and view all the answers

    What is one possible result when there are multiple precipitin lines observed?

    <p>Shows heterogeneity of the antibody for the antigen.</p> Signup and view all the answers

    What is one of the primary uses of counter-current immuno-electrophoresis?

    <p>Detection of antigens and antibodies in various body fluids</p> Signup and view all the answers

    Which of the following is a limitation of counter-current immuno-electrophoresis?

    <p>It is more expensive than agglutination-based tests</p> Signup and view all the answers

    What is the primary principle behind Radioimmunoassay (RIA)?

    <p>Antigen-antibody interaction involving radiolabeled antigens</p> Signup and view all the answers

    What type of isotopes are commonly used to label antigens in RIA?

    <p>Gamma-ray and beta-ray emitting isotopes</p> Signup and view all the answers

    In the context of RIA, what is the role of unlabeled antigens?

    <p>To compete with labeled antigens for antibody binding</p> Signup and view all the answers

    Which of the following advantages does counter-current immuno-electrophoresis have over double diffusion techniques?

    <p>It is faster and more sensitive</p> Signup and view all the answers

    What is a requirement for performing a Radioimmunoassay (RIA)?

    <p>Radiolabeled and unlabeled antigens</p> Signup and view all the answers

    Which of the following diseases is NOT commonly diagnosed using counter-current immuno-electrophoresis?

    <p>Tuberculosis</p> Signup and view all the answers

    What process is used to produce monoclonal antibodies?

    <p>Fusing B-lymphocytes with myeloma cells</p> Signup and view all the answers

    What is the main advantage of using chimeric monoclonal antibodies?

    <p>They have a longer serum half-life and reduced immunogenicity.</p> Signup and view all the answers

    Which type of monoclonal antibodies is considered the most common for cancer treatment?

    <p>Naked monoclonal antibodies</p> Signup and view all the answers

    How do conjugated monoclonal antibodies function in targeted therapy?

    <p>They transport drugs directly to the target antigens.</p> Signup and view all the answers

    Which of the following correctly describes humanized monoclonal antibodies?

    <p>Only the complementarity-determining regions are mouse-derived.</p> Signup and view all the answers

    What characteristics do murine monoclonal antibodies possess?

    <p>They have a high immunogenicity risk.</p> Signup and view all the answers

    What is a distinguishing feature of radiolabeled monoclonal antibodies?

    <p>They use a small radioactive particle attached to them.</p> Signup and view all the answers

    What is the role of monoclonal antibodies in targeting cancer cells?

    <p>They bind to unique sites on cancerous cells to target them.</p> Signup and view all the answers

    What is the primary purpose of attaching chemotherapy drugs to monoclonal antibodies?

    <p>To allow for targeted delivery of the drug to cancer cells</p> Signup and view all the answers

    Which animals are predominantly used in the initial production of monoclonal antibodies?

    <p>Laboratory mice</p> Signup and view all the answers

    What is the effect of the fusion between B-cells and myeloma cells in monoclonal antibody production?

    <p>It generates hybridoma cells capable of producing specific antibodies.</p> Signup and view all the answers

    Which of the following diseases is NOT commonly treated with monoclonal antibodies?

    <p>Diabetes mellitus</p> Signup and view all the answers

    What is a significant limitation of using mouse-derived monoclonal antibodies in human therapy?

    <p>They can cause the human immune system to generate antibodies against them.</p> Signup and view all the answers

    In the context of immunotherapy, what role do monoclonal antibodies play?

    <p>They enhance the body's immune response against specific diseases.</p> Signup and view all the answers

    Which technique is NOT used for the detection of diseases via monoclonal antibodies?

    <p>Radiotherapy</p> Signup and view all the answers

    What is the purpose of using polyethylene glycol in the production of monoclonal antibodies?

    <p>To facilitate the fusion of B-cells and myeloma cells</p> Signup and view all the answers

    Study Notes

    Immunoelectrophoresis

    • Procedure: A sample is diluted with protein diluent and applied to a gel. Electrophoresis is performed for 20 minutes at 100 volts. Antiserum is added to the gel and incubated for 18-20 hours. The gel is then washed and dried.
    • Results: The presence of elliptical precipitin lines indicates antigen-antibody interaction, while their absence suggests no reaction. Different proteins can be identified based on precipitation line intensity, shape, and position.
    • Applications: Used for identifying and quantifying proteins in serum, diagnosing monoclonal and polyclonal gammopathies, detecting normal and abnormal proteins, and analyzing complex protein mixtures.
    • Advantages: High resolving power, combining electrophoresis and immunodiffusion. Allows for identifying multiple antigens in serum.
    • Limitations: Slower, less sensitive, and more difficult to interpret than immunofixation electrophoresis. May not detect small monoclonal proteins. Limited availability of specific antibodies for food analysis.

    Counter-Current Immunoelectrophoresis

    • Procedure: Antigen mixture is separated by electrophoresis and then tested by immunodiffusion. Antigen and antibody move in opposite directions, forming precipitates where they meet in optimal concentrations.
    • Objectives: Rapidly check for the presence of antibodies for a specific antigen. Detect antigens and/or antibodies in serum for disease diagnosis.
    • Principle: Antigen moves towards the anode and antibody towards the cathode through agar under an electric field. Migration is facilitated, and precipitation lines are visible within 30-60 minutes.
    • Results: A precipitin line between antigen and antisera wells indicates a positive reaction or specific antigen-antibody interaction. Its absence indicates no reaction or absent antibody-antigen. Multiple precipitin lines indicate antibody heterogeneity for the antigen.
    • Applications: Rapid and specific detection of antigens and antibodies in serum, cerebrospinal fluid, and other body fluids. Diagnosing infectious diseases (bacterial, viral, fungal, parasitic). Detecting antigens like alpha-fetoprotein and capsular antigens. Used for Hepatitis B surface antigen (HBsAg), fetoprotein, hydatid and amoebic antigens, cryptococcal antigen, and pneumococcal capsular antigens.
    • Advantages: Fast antigen-antibody detection (30 minutes). More sensitive than electro-immunodiffusion.
    • Limitations: More expensive than agglutination tests. Decreased sensitivity, speed, and simplicity compared to other methods.

    Radioimmunoassay (RIA)

    • Principle: Antigen-antibody interaction with high sensitivity using radioisotopes.
    • Requirements: Radiolabeled antigens (eg., I-125, Tritium). Specific antibodies. Unlabeled antigens. Microtitre plates. Washing buffer.
    • Procedure: Radiolabeled antigens compete with unlabeled antigens from the sample to bind to specific antibodies. Unlabeled antigens displace labeled antigens.
    • Applications: Determination of antigens or antibodies in a sample.

    Monoclonal Antibodies

    • Definition: Antibodies produced from a single clone of cells.
    • Production: Immunization of an animal with a specific antigen. B-cells from the animal's spleen are fused with myeloma cells. The resulting hybridomas produce monoclonal antibodies.
    • Types based on origin: Murine, Chimeric, Humanized, Human.
    • Types based on function: Naked, Conjugated (Radiolabeled, Chemolabeled), Bispecific
    • Applications: Treatment of diverse diseases and disorders (cancers, autoimmune diseases, inflammatory conditions). Diagnostic tools to detect circulating antigens and antibodies.
    • Limitations: Immunogenicity in patients (serum sickness). Potential for antibody production against mouse antibodies.

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    Description

    This quiz covers the essential aspects of immunoelectrophoresis, including its procedure, results interpretation, applications, advantages, and limitations. Test your knowledge on how this technique is used to identify and quantify proteins in serum and analyze complex protein mixtures.

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