20 Questions
What is the primary focus of Immuno assays?
Isolating or quantifying the level of specific proteins
What is the main purpose of Immunoblotting?
Looking at contents of a specific protein of interest
What is the function of Immunoprecipitation?
Using antibodies to recognize specific proteins like T4, insulin, glucagon, etc.
What is the primary purpose of FACS (Fluorescence-activated cell sorting)?
Quantifying the level of specific proteins
In what context are antibodies used against the EGF receptor?
Therapeutic purposes
Which technique is used to detect the amount of a specific protein in a mixture?
Immunoblotting
What is the purpose of incubating cells with antibodies that recognize cell surface proteins in Fluorescence Assisted Cell Sorting (FACS)?
To sort cells based on molecular characteristics
What is the main principle behind immunoprecipitation?
Using antibodies to purify an antigen present in a complex mixture
What is the function of charged deflector plates in Fluorescence Assisted Cell Sorting (FACS)?
To separate various cell populations present in a mixture
What is the role of secondary antibody conjugated at the surface of beads in immunoprecipitation?
To pull down and separate antibody from the mixture
In competitive ELISA, what is the purpose of using an inhibitor antigen?
To compete with the target protein for antibody binding
What is the primary advantage of sandwich ELISA over direct and indirect ELISAs?
It is more specific as the antigen has to bind to two antibodies to be detected
What is the purpose of epitope mapping in antibody design and vaccine development?
To experimentally identify the binding sites of antibodies on their target antigen
What is the main difference between direct labeling and indirect labeling in immuno-microscopy?
Direct labeling involves one antibody as both the primary and detector, while indirect labeling uses two antibodies
What enables ELISAs to be quantitative when used with a standard curve?
Known amounts of antigen are used for comparison
Which statement about polyclonal antibodies is most accurate?
Polyclonal antibodies are a population of antibodies binding different epitopes of the same antigen, generated by different B cell clones and primarily used for research and diagnosis.
How are monoclonal antibodies generated?
Monoclonal antibodies are generated by a single B-cell clone called hybridoma and are used in various fields including therapy due to their specificity and reproducibility.
Which statement best describes the use of monoclonal antibodies?
Monoclonal antibodies are used in therapy due to their specificity and reproducibility between batches, and they can be tested for reactivity to the antigen and grown in large quantities.
What is the major difference between polyclonal and monoclonal antibodies?
Polyclonal antibodies are a population of antibodies binding different epitopes of the same antigen, while monoclonal antibodies bind a single epitope on the antigen.
Which type of antibodies are primarily used for research and diagnosis?
Polyclonal antibodies
Study Notes
Harnessing the Immune System for Antibody Production
- Antibodies are produced in response to antigens and are naturally produced by B-lymphocytes in the body's adaptive immune system.
- Antibodies are composed of two heavy chains and two light chains, with a variable region containing the specific antigen binding site and a constant region divided into five different classes.
- The two major classes of antibodies used in immunochemistry are polyclonal and monoclonal antibodies, each with distinct characteristics and applications.
- Polyclonal antibodies are a population of antibodies binding different epitopes of the same antigen, generated by different B cell clones and primarily used for research and diagnosis.
- Polyclonal antibodies are produced in animals by injecting them with antigens, isolating and purifying the antibodies from the animal blood serum.
- Monoclonal antibodies bind a single epitope on the antigen, are generated by a single B-cell clone called hybridoma, and are used in various fields including therapy due to their specificity and reproducibility.
- Hybridomas are obtained by fusing single lymphocytes from immunized mice with a single cancer cell, and the clone producing the antibody with the best properties is selected and grown in large quantities.
- Antibodies can be conjugated with various molecules to allow detection by colorimetric and fluorimetric methods, immuno selection, immuno precipitation, and immunoassays.
- The vast majority of immunoassays, particularly enzyme immunosorbent assays (ELISA), are commonly used for diagnosing infectious agents such as viruses and quantitatively measuring substances in blood like hormones, cytokines, and chemokines in research and hospital settings.
- Polyclonal antibodies are cheaper to produce but less specific and more variable from batch to batch, while monoclonal antibodies are more expensive to produce but have less batch to batch variation and are more specific.
- Animals like rabbits, goats, and donkeys are usually employed to produce polyclonal antibodies due to their larger size, while mice and rats are not used as they are too small.
- Monoclonal antibodies are used in therapy due to their specificity and reproducibility between batches, and they can be tested for reactivity to the antigen and grown in large quantities.
Test your knowledge of antibody production and immune system harnessing with this quiz. Explore the differences between polyclonal and monoclonal antibodies, their applications, and the process of antibody production. Dive into the world of immunochemistry and learn about the various techniques and assays used in research and diagnostics.
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