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Questions and Answers
What is the first step in preparing a sample for immunoelectrophoresis?
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?
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?
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?
What is an application of immunoelectrophoresis in clinical diagnostics?
What is the purpose of staining the gel during immunoelectrophoresis?
What is the purpose of staining the gel during immunoelectrophoresis?
What characteristic is used to identify different proteins in immunoelectrophoresis?
What characteristic is used to identify different proteins in immunoelectrophoresis?
Which condition might immunoelectrophoresis help diagnose?
Which condition might immunoelectrophoresis help diagnose?
What is one of the advantages of using immunoelectrophoresis?
What is one of the advantages of using immunoelectrophoresis?
What is one major limitation of immunoelectrophoresis compared to immunofixation electrophoresis?
What is one major limitation of immunoelectrophoresis compared to immunofixation electrophoresis?
What does counter current immunoelectrophoresis require for the movement of antigen and antibody?
What does counter current immunoelectrophoresis require for the movement of antigen and antibody?
What indicates a positive reaction in counter current immunoelectrophoresis?
What indicates a positive reaction in counter current immunoelectrophoresis?
What is a common application of counter current immunoelectrophoresis?
What is a common application of counter current immunoelectrophoresis?
In counter current immunoelectrophoresis, where do the antigen and antibody migrate?
In counter current immunoelectrophoresis, where do the antigen and antibody migrate?
What does the absence of a precipitin line in counter current immunoelectrophoresis indicate?
What does the absence of a precipitin line in counter current immunoelectrophoresis indicate?
What is a potential drawback of using immunoelectrophoresis in food analysis?
What is a potential drawback of using immunoelectrophoresis in food analysis?
What is one possible result when there are multiple precipitin lines observed?
What is one possible result when there are multiple precipitin lines observed?
What is one of the primary uses of counter-current immuno-electrophoresis?
What is one of the primary uses of counter-current immuno-electrophoresis?
Which of the following is a limitation of counter-current immuno-electrophoresis?
Which of the following is a limitation of counter-current immuno-electrophoresis?
What is the primary principle behind Radioimmunoassay (RIA)?
What is the primary principle behind Radioimmunoassay (RIA)?
What type of isotopes are commonly used to label antigens in RIA?
What type of isotopes are commonly used to label antigens in RIA?
In the context of RIA, what is the role of unlabeled antigens?
In the context of RIA, what is the role of unlabeled antigens?
Which of the following advantages does counter-current immuno-electrophoresis have over double diffusion techniques?
Which of the following advantages does counter-current immuno-electrophoresis have over double diffusion techniques?
What is a requirement for performing a Radioimmunoassay (RIA)?
What is a requirement for performing a Radioimmunoassay (RIA)?
Which of the following diseases is NOT commonly diagnosed using counter-current immuno-electrophoresis?
Which of the following diseases is NOT commonly diagnosed using counter-current immuno-electrophoresis?
What process is used to produce monoclonal antibodies?
What process is used to produce monoclonal antibodies?
What is the main advantage of using chimeric monoclonal antibodies?
What is the main advantage of using chimeric monoclonal antibodies?
Which type of monoclonal antibodies is considered the most common for cancer treatment?
Which type of monoclonal antibodies is considered the most common for cancer treatment?
How do conjugated monoclonal antibodies function in targeted therapy?
How do conjugated monoclonal antibodies function in targeted therapy?
Which of the following correctly describes humanized monoclonal antibodies?
Which of the following correctly describes humanized monoclonal antibodies?
What characteristics do murine monoclonal antibodies possess?
What characteristics do murine monoclonal antibodies possess?
What is a distinguishing feature of radiolabeled monoclonal antibodies?
What is a distinguishing feature of radiolabeled monoclonal antibodies?
What is the role of monoclonal antibodies in targeting cancer cells?
What is the role of monoclonal antibodies in targeting cancer cells?
What is the primary purpose of attaching chemotherapy drugs to monoclonal antibodies?
What is the primary purpose of attaching chemotherapy drugs to monoclonal antibodies?
Which animals are predominantly used in the initial production of monoclonal antibodies?
Which animals are predominantly used in the initial production of monoclonal antibodies?
What is the effect of the fusion between B-cells and myeloma cells in monoclonal antibody production?
What is the effect of the fusion between B-cells and myeloma cells in monoclonal antibody production?
Which of the following diseases is NOT commonly treated with monoclonal antibodies?
Which of the following diseases is NOT commonly treated with monoclonal antibodies?
What is a significant limitation of using mouse-derived monoclonal antibodies in human therapy?
What is a significant limitation of using mouse-derived monoclonal antibodies in human therapy?
In the context of immunotherapy, what role do monoclonal antibodies play?
In the context of immunotherapy, what role do monoclonal antibodies play?
Which technique is NOT used for the detection of diseases via monoclonal antibodies?
Which technique is NOT used for the detection of diseases via monoclonal antibodies?
What is the purpose of using polyethylene glycol in the production of monoclonal antibodies?
What is the purpose of using polyethylene glycol in the production of monoclonal antibodies?
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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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