Podcast
Questions and Answers
Which immunoglobulin is primarily associated with seasonal allergies to tree pollen?
Which immunoglobulin is primarily associated with seasonal allergies to tree pollen?
What is the molecular weight of the Fab fragments produced by papain digestion of IgG?
What is the molecular weight of the Fab fragments produced by papain digestion of IgG?
Which antibody fragment is known for having a lower tendency to bind non-specifically?
Which antibody fragment is known for having a lower tendency to bind non-specifically?
What role does the Fc region of IgG play in immunoaffinity chromatography?
What role does the Fc region of IgG play in immunoaffinity chromatography?
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Which of the following statements about F(ab')2 fragments is true?
Which of the following statements about F(ab')2 fragments is true?
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Which class of antibody is the major component of the secondary immune response and makes up 70-75% of normal serum?
Which class of antibody is the major component of the secondary immune response and makes up 70-75% of normal serum?
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What percentage of the immunoglobulin pool does IgM represent?
What percentage of the immunoglobulin pool does IgM represent?
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Which antibody class is primarily found in mucous, saliva, tears, and breast milk?
Which antibody class is primarily found in mucous, saliva, tears, and breast milk?
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Which class of antibodies is mainly involved in activating basophils and mast cells?
Which class of antibodies is mainly involved in activating basophils and mast cells?
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Which antibody class is known to respond to parasitic worm infections and is associated with allergic reactions?
Which antibody class is known to respond to parasitic worm infections and is associated with allergic reactions?
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What is the primary role of IgM in the immune response?
What is the primary role of IgM in the immune response?
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Which antibody class is primarily secreted by plasma cells into the bloodstream and can cross the placenta?
Which antibody class is primarily secreted by plasma cells into the bloodstream and can cross the placenta?
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What is the role of IgA in the immune system?
What is the role of IgA in the immune system?
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Study Notes
Introduction to Immunoassays
- This lecture introduces immunoanalysis, a key aspect of biological research and diagnostics.
- The lecture uses IgG as a visual example, showing its structure.
Learning Objectives
- Students will be able to list different antibody (Ab) classes.
- Students will be able to describe the function of different Ab classes.
- Students will identify the components of an antibody.
- Students will understand how antibodies (IgG) are broken down by digestion enzymes.
- Key components of an antibody include: VL and CL (light chains); VH and CH (heavy chains); Fab; Fc; Hinge region
Immune Cells
- Immune cells originate from a hematopoietic stem cell (Hemocytobast).
- Types of immune cells include lymphocytes, monocytes, eosinophils, basophils, neutrophils, thrombocytes and erythrocytes.
- Lymphocytes are white blood cells.
Antibody Classes
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IgG:
- Major immunoglobulin (Ig) in normal serum (70-75%).
- A key component in secondary immune response.
- Plays a role in immobilizing and agglutinating pathogens for phagocytosis.
- Single immunoglobulin molecule.
-
IgM:
- Accounts for approximately 10% of the Ig pool.
- A primary immune response antibody the first Ig in the human fetus.
- Abundant in responses against complex antigens.
- Activates complement.
-
IgA:
- Accounts for approximately 15 to 20% of plasma Ig types.
- Predominant Ig in mucosal secretions (e.g., saliva, milk).
- Protects against respiratory and gastrointestinal infections.
-
IgD:
- Present in very small amounts in the human body pool.
- Found on immature B cells.
- Enhances mucosal homeostasis and immune surveillance.
-
IgE:
- Commonly associated with allergic diseases (e.g., asthma).
- Important in responses to parasitic infections.
- Stimulates eosinophils and histamine release from mast cells and basophils.
Group Activity – Antibody Selection
- This activity requires choosing the appropriate antibody type for specific immune responses (e.g., Respiratory illness, seasonal allergies).
Types of Antibodies (Immunoglobulins)
- Antibodies (Ig) are important for targeting antigens.
- IgG, IgA, IgM have different functions within the body.
- Detailed information on different antibody's types and specific functions is included.
IgG - Predominantly used in immunoassays
- IgG has two light chains and two heavy chains.
- The structure includes variable regions (VL, VH) and constant regions (CL, CH)
- The hinge region of the antibody provides flexibility, important for binding to antigens.
- Fab and Fc are fragments of the antibody molecule.
Digestion of Antibody Fragments
- Papain cleavage of IgG into Fab and Fc fragments.
- Pepsin reaction resulting in F(ab')2 and Fc fragments.
- Fragment sizes and molecular weights are reported.
Ab Fragment Applications
- Antibodies (Abs) are applied in immunoassays
- The Ab Fc region is anchored for enhanced antigen binding to exposed Fab regions.
- Methods like direct, indirect, sandwich, and competitive ELISA are used in immunoassays.
- Fragment applications in different research areas like chromatin immunoprecipitation (CHIP).
- Applications in areas including DNA purification and analysis.
Ab Fragment Applications – Antibody Fragments
- Fab regions are smaller sized, less likely to bind nonspecifically.
- Applications include immunoaffinity chromatography, purification of IgG.
- Use in various molecular biology techniques including DNA purification and analysis.
Advantages of Antibodies in Immunoassays
- Advantages of applying antibodies, including stability, abundance, ease of preparation, and modifiability.
- Applicability to specific situations, including antibody mixing with related specificities (polyclonal) or single binding specificity (monoclonal). - Binding to antigens (Ag) and non-antigenic molecules (haptens).
Abs Fragment Applications - Novel applications
- Novel diagnostic and therapeutic developments are possible, involving small antibody fragments.
- Fragment sizes as small as 3kDa.
- Antibody fragments can be labelled with radioisotopes or fluorescent molecules to target specific tissues (e.g., tumours)
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Description
This quiz covers the key concepts of immunoassays, focusing on antibodies, especially IgG, and their functions in biological research and diagnostics. Students will explore antibody classes, immune cells, and the components of antibodies. Prepare to test your knowledge on the fundamentals of immunology.
