Antibody-antigen interactions, ELISA

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Life Science
LIFE 212
Farida Safadi- Chamberlain

29 August Lab 2 Pre-Lab Write-Up Theory (or background information) of the use of ELISA (Enzyme-Linked Immunosorbent Assays) for detection of protein antigens Scientific question to be answered Hypothesis and predicted results Objective of this experiment Antibody – Antigen (protein) Interactions A special type of Protein – Protein Interaction Antibodies are a type of immune-response protein made in vertebrates Used to detect disease pathogens Is used in many applications in research labs Antibodies are proteins that are produced as part of the adaptive immune response to potential pathogens – foreign antigens Characteristics of Antibodies (Abs) Abs are produced on the surface of white blood cells (B cells): Many types of Ab molecules are produced at different times in the immune response Part of the immunoglobin protein class Can be Monoclonal Polyclonal Antigens are defined as any foreign substance detected by the immune system Each Ab binds to a single Ag binding site (epitope) Response to a single antigen results in many types of antibodies (polyclonal antibodies) Many Assays Use Monoclonal Abs Monoclonal Abs are produced from hybridoma cells They recognize a single antigenic epitope Lab 2 Use an immunoassay to detect the antigen-antibody response Immunoassays are highly SPECIFIC and SENSITIVE Specificity is based on the highly precise interaction between antibodies and their antigens (Ab/Ag) Sensitivity is based on the ability to detect a very few molecules in a complex mixture. Therefore, high sensitivity is based on a low detection
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