20.1 Polyclonal and Monoclonal Antibody Production
3 min read•june 18, 2024
are powerful tools in immunology, with diverse applications in diagnostics and therapeutics. recognize multiple , while target specific ones. Understanding their production methods and characteristics is crucial for their effective use.
have revolutionized medicine by reducing immunogenicity and improving compatibility with the human immune system. These engineered antibodies are used in cancer immunotherapy, autoimmune disease treatment, and infectious disease prevention, showcasing their versatility in modern healthcare.
Antibody Production and Applications
Monoclonal vs polyclonal antibodies
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-
- Produced by multiple in response to an
- Recognize multiple on the same
- Production methods involve immunizing an animal (, , or ) with the antigen and collecting containing the antibodies
- Applications include , ELISA (Enzyme-Linked Immunosorbent Assay), and
- Monoclonal antibodies
- Produced by a single B cell clone recognizing a specific epitope on an antigen
- Production methods use by immunizing a with the antigen, fusing B cells from the mouse's with , and selecting and culturing the that produce the desired antibody
- Applications include diagnostic tests (pregnancy tests, cancer biomarker detection), targeted cancer therapies (, ), and treatment of autoimmune diseases ( for rheumatoid arthritis)
Antibody cross-reactivity in diagnostics
- occurs when an antibody recognizes and binds to an epitope on an antigen other than its intended target due to structural similarities between epitopes on different antigens
- Implications for diagnostic testing include
- results where cross-reactive antibodies bind to non-target antigens, leading to incorrect positive test results and potentially causing unnecessary treatment or further testing
- results where cross-reactive antibodies bind to the target antigen with lower , reducing the sensitivity of the test and potentially leading to missed diagnoses and delayed treatment
- Reduced (ability to correctly identify individuals without the condition) and sensitivity (ability to correctly identify individuals with the condition) of diagnostic tests
Production of humanized monoclonal antibodies
- are engineered to contain more human protein sequences to reduce immunogenicity and improve compatibility with the human immune system
- Production process involves
- Identifying the () of a mouse monoclonal antibody
- Grafting the CDRs onto a human antibody framework
- Expressing the humanized antibody in a mammalian cell line
- Purifying and characterizing the humanized antibody
- Clinical uses include
- Cancer immunotherapy by targeting specific tumor antigens (, ) and enhancing the immune system's ability to recognize and destroy cancer cells
- Treatment of inflammatory and autoimmune diseases by neutralizing pro-inflammatory cytokines (, ) and modulating the immune response to reduce tissue damage
- Infectious disease prevention and treatment by neutralizing viral or bacterial pathogens (, ) and providing passive immunity in high-risk individuals
Antibody characteristics and production techniques
- Antigen: A substance that stimulates an immune response, typically foreign molecules like proteins or polysaccharides
- : The process of exposing an animal to an antigen to stimulate antibody production
- : The strength of binding between an antibody and its specific antigen
- : The ability of an antibody to bind only to its intended target antigen
- : A hybrid cell produced by fusing a B cell with a tumor cell, used in monoclonal antibody production
- : The technique used to create hybridomas by combining B cells with myeloma cells
Key Terms to Review (65)
Adjuvant: An adjuvant is a substance that enhances the body's immune response to an antigen. It is often used in vaccines to improve their efficacy.
Affinity: Affinity refers to the strength of the interaction between an antibody and its specific antigen. It is a measure of how tightly an antibody binds to its antigenic target.
Affinity: Affinity refers to the strength of the binding interaction between an antibody and its target antigen. It is a measure of how tightly an antibody can bind to a specific epitope on an antigen, which is crucial for the antibody's ability to recognize and neutralize the target effectively.
Anthrax Toxin: Anthrax toxin is a potent virulence factor produced by the bacterium Bacillus anthracis, the causative agent of the deadly disease anthrax. This complex toxin plays a crucial role in the pathogenesis of anthrax and is a key target for understanding bacterial virulence and the development of countermeasures.
Antibodies: Antibodies are specialized proteins produced by B cells that recognize and neutralize specific antigens. They play a crucial role in the adaptive immune response by identifying and binding to foreign pathogens.
Antigen: An antigen is any substance that induces an immune response in the body, specifically by triggering the production of antibodies. They are typically foreign proteins or polysaccharides found on the surface of pathogens like bacteria and viruses.
Antigen: An antigen is a substance, typically a protein or polysaccharide, that is capable of stimulating an immune response and triggering the production of antibodies. Antigens are central to the topics of polyclonal and monoclonal antibody production, as well as enzyme-linked immunosorbent assays (ELISAs) and enzyme immunoassays (EIAs).
Antiserum: Antiserum is a blood serum containing polyclonal antibodies that can neutralize specific antigens, such as pathogens or toxins. It is used in immunotherapy and diagnostic tests to confer passive immunity or identify substances.
Avidity: Avidity refers to the overall strength of binding between an antibody and its antigen, involving multiple binding sites. It is a cumulative measure that takes into account both the affinity of each individual interaction and the valency of the antibody.
B Cell Clones: B cell clones refer to the identical copies of a single B lymphocyte that are generated through rapid cell division in response to an antigen. These clones produce large quantities of antibodies specific to the triggering antigen, providing a targeted immune response.
CDRs: CDRs, or Complementarity Determining Regions, are the most variable parts of an antibody molecule that are responsible for recognizing and binding to specific antigens. These regions are critical for the immune system's ability to generate a diverse repertoire of antibodies that can recognize a wide range of foreign substances.
Cell Fusion: Cell fusion is the process by which two or more cells merge their membranes and contents, forming a single, unified cell. This phenomenon is crucial in various biological contexts, including polyclonal and monoclonal antibody production.
Complementarity-Determining Regions: Complementarity-determining regions (CDRs) are the hypervariable regions within the variable domains of antibody heavy and light chains that are responsible for antigen binding. These regions are the most diverse parts of an antibody and play a crucial role in the immune system's ability to recognize a wide variety of antigens.
Confirmatory test: A confirmatory test is a secondary assay performed to validate the results obtained from an initial screening test, ensuring accuracy and specificity. It is essential in eliminating false positives and confirming the presence of specific antibodies or antigens.
Cross-reactivity: Cross-reactivity occurs when an antibody directed against one specific antigen also reacts with a different, but structurally similar, antigen. This phenomenon can have significant implications in diagnostic tests and immune responses.
Cross-reactivity: Cross-reactivity refers to the ability of an antibody or antigen to bind to multiple, structurally similar targets, leading to potential false-positive or false-negative results in various immunological assays and diagnostic tests.
CTLA-4: CTLA-4, or Cytotoxic T-Lymphocyte-Associated Protein 4, is a crucial immune checkpoint receptor that plays a pivotal role in regulating the activation and function of T cells, a key component of the adaptive immune system. This term is particularly relevant in the context of cancer immunobiology and immunotherapy, as well as in the production of polyclonal and monoclonal antibodies.
Epitopes: Epitopes are specific regions on an antigen that are recognized and bound by antibodies or T-cell receptors. They play a crucial role in the specificity of the immune response.
Epitopes: Epitopes, also known as antigenic determinants, are the specific regions on the surface of an antigen that are recognized and bound by antibodies or T cell receptors. These unique molecular structures on the surface of pathogens or foreign substances are crucial for the activation of the adaptive immune response.
False-negative: A false-negative result occurs when a test incorrectly indicates the absence of a condition or substance. This can lead to missed diagnoses and delayed treatments.
False-Negative: A false-negative result occurs when a test or diagnostic procedure fails to detect the presence of a condition or disease that is actually present. This can lead to a delay in treatment or a missed diagnosis, which can have significant consequences for the individual's health.
False-positive: A false-positive occurs when a test incorrectly indicates the presence of a condition, such as an infection or disease, when it is not actually present. This can lead to misdiagnosis and unnecessary treatment.
False-Positive: A false-positive is a test result that incorrectly indicates the presence of a condition or characteristic when it is actually not present. This term is particularly relevant in the context of polyclonal and monoclonal antibody production, where it can impact the accuracy and reliability of immunoassays and diagnostic tests.
Goat: A goat is a domesticated ruminant mammal that is raised for its milk, meat, and fiber. Goats are known for their agility, hardiness, and ability to thrive in a variety of environments, making them an important livestock animal in many parts of the world.
Hepatitis C: Hepatitis C is a liver infection caused by the hepatitis C virus (HCV). It can lead to both acute and chronic hepatitis, ranging in severity from a mild illness to a serious, lifelong condition.
Horse: A horse is a large, hoofed mammal that has been domesticated and used by humans for transportation, work, and recreation for thousands of years. In the context of polyclonal and monoclonal antibody production, horses are often used as a source of antibodies due to their ability to generate a diverse range of immune responses.
Humanized monoclonal antibodies: Humanized monoclonal antibodies are engineered antibodies from non-human species whose protein sequences have been modified to increase their similarity to antibodies produced naturally in humans. This reduces the immune response against them when used therapeutically.
Humanized Monoclonal Antibodies: Humanized monoclonal antibodies are a type of therapeutic antibody that have been engineered to reduce their immunogenicity by replacing most of the non-human (typically mouse) components with human counterparts. This allows for more effective and safer use in human patients compared to fully non-human monoclonal antibodies.
Hybridoma: A hybridoma is a cell line created by fusing an antibody-producing B-cell with a myeloma (cancer) cell, resulting in a hybrid cell that can produce large quantities of monoclonal antibodies. These cells are used extensively in research and diagnostic applications due to their ability to produce identical antibodies indefinitely.
Hybridoma: A hybridoma is a hybrid cell line created by fusing an antibody-producing B lymphocyte with a myeloma (cancer) cell. This fusion allows for the continuous production of a specific monoclonal antibody, making it a crucial tool in the context of polyclonal and monoclonal antibody production.
Hybridoma Cells: Hybridoma cells are a type of immortalized cell line created by fusing an antibody-producing B lymphocyte with a myeloma (cancer) cell. This fusion results in a hybrid cell that can continuously produce a single, specific monoclonal antibody, making it a powerful tool for antibody production and research.
Hybridoma Technology: Hybridoma technology is a method used to produce large quantities of highly specific monoclonal antibodies. It involves fusing an antibody-producing B cell with an immortal myeloma cell, creating a hybrid cell line called a hybridoma that can continuously secrete the desired antibody.
IL-6: IL-6 (Interleukin-6) is a multifunctional cytokine that plays a crucial role in the immune response, inflammation, and the regulation of various physiological processes. It is a key player in the context of polyclonal and monoclonal antibody production, as it influences the differentiation and activation of B cells and plasma cells.
Immunization: Immunization is the process of inducing immunity to an infectious disease by administering a vaccine. It helps the body develop the necessary antibodies and immune responses to protect against future exposure to that disease.
Immunohistochemistry: Immunohistochemistry (IHC) is a powerful technique that combines immunology and histology to detect and visualize specific proteins or antigens within cells and tissues. It is widely used in both research and clinical settings to study the distribution and localization of target molecules in biological samples.
Immunohistochemistry (IHC): Immunohistochemistry (IHC) is a laboratory technique used to visualize the presence and localization of specific antigens in tissue sections by using labeled antibodies. It combines histological, immunological, and biochemical techniques to identify cells or tissue components.
In vitro assays: In vitro assays are experimental procedures conducted in a controlled environment outside of a living organism, often in petri dishes or test tubes. They are used to study biological processes, including the immune response.
Infliximab: Infliximab is a monoclonal antibody used as a therapeutic agent in the treatment of various inflammatory and autoimmune conditions. It is a key term in the context of understanding polyclonal and monoclonal antibody production, as it represents an example of a clinically important monoclonal antibody drug.
Monoclonal Antibodies: Monoclonal antibodies are identical antibodies produced by a single clone of B cells, each recognizing a specific epitope on an antigen. They are a crucial tool in immunology, diagnostics, and therapeutic applications, with their ability to target specific molecules with high affinity and specificity.
Monoclonal antibodies (mAbs): Monoclonal antibodies (mAbs) are laboratory-produced molecules engineered to bind to specific antigens. They are used extensively in diagnostics, therapeutics, and research.
Mouse: A mouse is a small rodent that is commonly used in laboratory research, particularly in the production and study of polyclonal and monoclonal antibodies. Mice are widely employed as model organisms due to their genetic similarity to humans, ease of handling, and well-established experimental protocols.
Myeloma Cells: Myeloma cells are abnormal plasma cells that originate from B lymphocytes and proliferate uncontrollably, leading to the development of multiple myeloma, a type of blood cancer. These malignant cells produce large quantities of a single type of antibody, which can be detected in the blood or urine of affected individuals.
PD-1: PD-1, or Programmed Cell Death Protein 1, is an inhibitory receptor expressed on the surface of T cells, B cells, and other immune cells. It plays a crucial role in regulating the immune response and maintaining self-tolerance, making it a key player in both cancer immunobiology and immunotherapy.
Plantibodies: Plantibodies are antibodies produced in genetically modified plants. They are used for therapeutic and diagnostic purposes due to their ability to be mass-produced efficiently.
Polyclonal antibodies: Polyclonal antibodies are a mixture of antibodies produced by different B cell clones in the body, each recognizing a different epitope on the same antigen. They are commonly used in research and diagnostic laboratories to detect antigens due to their high sensitivity.
Polyclonal Antibodies: Polyclonal antibodies are a diverse mixture of antibodies produced by multiple B-cell lineages in response to an antigen. They recognize multiple epitopes on the same antigen and are commonly used in various immunological techniques, such as those covered in Chapters 20.1 and 20.5.
Polyclonal antibody: Polyclonal antibodies are a mixture of antibodies produced by different B cell clones in the body, each recognizing a different epitope on the same antigen. They are commonly used in research and diagnostic applications due to their ability to recognize multiple epitopes.
Portal specificity: Portal specificity refers to the phenomenon where certain pathogens cause disease only when they enter the host through specific anatomical sites. This specificity is due to factors such as receptor availability, environmental conditions, and immune response at the entry site.
Rabbit: A rabbit is a small mammal of the order Lagomorpha, characterized by long ears, a short fluffy tail, and strong hind legs. In the context of 20.1 Polyclonal and Monoclonal Antibody Production, the rabbit is a commonly used animal model for the generation of antibodies.
Respiratory syncytial virus: Respiratory Syncytial Virus (RSV) is a common respiratory virus that causes infections of the lungs and respiratory tract. It is particularly dangerous for infants, elderly adults, and immunocompromised individuals.
Respiratory Syncytial Virus: Respiratory syncytial virus (RSV) is a highly contagious virus that primarily infects the respiratory tract, causing a range of illnesses from mild cold-like symptoms to severe pneumonia, especially in young children and older adults. This virus is a major focus in the context of polyclonal and monoclonal antibody production, as well as viral infections of the respiratory tract.
Rituximab: Rituximab is a monoclonal antibody used to target CD20 proteins on the surface of B cells. It is often used in the treatment of certain autoimmune diseases and lymphomas.
Rituximab: Rituximab is a monoclonal antibody used in the treatment of various types of cancer and autoimmune disorders. It specifically targets the CD20 antigen expressed on the surface of B cells, making it a crucial component of cancer immunobiology and immunotherapy approaches.
Screening tests: Screening tests are preliminary assessments used to detect the presence of specific antibodies or antigens in a sample. They are crucial for identifying immune responses and guiding further diagnostic testing.
Serum: Serum is the clear, yellowish liquid component of blood that remains after the removal of blood cells and clotting proteins. It is a critical component in the production and analysis of antibodies, playing a central role in polyclonal and monoclonal antibody production.
Specificity: Specificity in immunology refers to the ability of the adaptive immune system to target specific antigens. This ensures that immune responses are directed against particular pathogens, minimizing collateral damage to host tissues.
Specificity: Specificity refers to the ability of a biological entity, such as an antibody or receptor, to recognize and bind to a particular target or antigen with a high degree of precision and selectivity. This term is crucial in understanding the adaptive immune response and the production of monoclonal and polyclonal antibodies.
Spleen: The spleen is an organ located in the upper left abdomen that filters blood, removes old red blood cells, and plays a critical role in immune response by producing white blood cells.
Spleen: The spleen is an organ located in the upper left part of the abdomen, behind the stomach. It plays a crucial role in the body's immune response and the filtration of blood, serving as an important component of both the circulatory and lymphatic systems.
Test sensitivity: Test sensitivity measures the proportion of actual positives correctly identified by a diagnostic test. High sensitivity means fewer false negatives.
Test specificity: Test specificity is the ability of a diagnostic test to correctly identify those without the disease (true negative rate). It measures how well the test can distinguish between the target analyte and non-target substances.
TNF-α: TNF-α, or Tumor Necrosis Factor-alpha, is a pro-inflammatory cytokine produced primarily by activated macrophages and other immune cells. It plays a crucial role in the regulation of immune responses, inflammation, and cell death.
Trastuzumab: Trastuzumab is a monoclonal antibody that targets the HER2 protein, which is overexpressed in certain types of breast cancer. It is a key therapeutic agent used in the treatment of HER2-positive breast cancer, a particularly aggressive form of the disease.
Variable regions: Variable regions are the parts of an antibody's structure that vary between different antibodies and are responsible for antigen binding. These regions determine the specificity and diversity of the immune response.
Western Blotting: Western blotting is an analytical technique used to detect and quantify specific proteins in a complex mixture of proteins extracted from cells or tissues. It involves separating proteins by size through gel electrophoresis, transferring them to a membrane, and using antibodies to identify and visualize the target proteins.