Antibody-Antigen - Basis and Handling/Storage Conditions
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Course
AU858 (BIOL07013)
Institution
ATU Sligo
These notes over week 1 and 2 of Immunodiagnostics. Which are the basic structures and overview of the immunoglobulins and their purpose. As well the required storage conditions for long term storage. Easy read and understand. Highlighted important parts for ease
Table of Contents
1. Immunoglobulin – structure and definition
2. Classes of Immunoglobulins
3. Polyclonal
4. Monoclonal
5. Hapten's and Hybridoma's
6. Epitope and Antigens
7. Characteristics of good antigens
8. Purification Techniques
9. Chimeric and Humanized
10. Primary antibodies and Secondary antibodies
11. Affinity, Avidity and Specificity
12. Crosslinking and Bioconjugation
13. Molecules added to Antibodies
14. Storage
Antibodies
Definition and Structure
• Antibodies are immunoglobulins that bind to specific antigens, produced in response to
foreign molecules in the body.
•
• Image: The University of Arizona (2019). Antibody Structure. [online] Arizona.edu. Available at:
http://www.biology.arizona.edu/immunology/tutorials/antibody/structure.html.
• They consist of Y-shaped units with heavy and light chains, where light chains include
variable (VL) and constant (CL) domains.
• Fc region: Is glycosylated, has many effector functions, serves to distinguish
one class of ab from another
•
Antibody Domains and Fragments
• Heavy and light chains contain immunoglobulin domains with variable (VH) and constant
regions.
• Enzymatic cleavage of IgG yields F(ab')2 and Fab fragments, with distinct antigen-binding
regions.
Isotypes and Classes
• Antibodies are classified into IgG, IgM, IgA, IgD, and IgE based on structure and function.
MADGE
• IgG antibodies further divide into subclasses (IgG1, IgG2, IgG3, IgG4) with varying disulfide
bonds and hinge region lengths.
• IgG is the biggest percentage of immunoglobulins at 80%, it is passed through the
placenta from the mother
• IgE is the first produced at an allergic reaction
Production
Polyclonal Antibodies
• Polyclonal antibodies are collected from multiple cells, commonly produced in animals
like rabbits.
• Production involves immunization, regular test bleeds, and cardiac bleed for high-titer
antiserum.
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