Monoclonal and polyclonal antibodies are produced through different processes. Monoclonal antibodies are derived from a single clone and recognize a single epitope, while polyclonal antibodies recognize multiple epitopes. Monoclonal antibodies are produced through hybridoma technology involving immunizing an animal, fusing lymphocytes with myeloma cells, and screening for antibody-producing clones. Polyclonal antibodies involve immunizing an animal and purifying the antibody response. Both have applications in research, diagnosis, and therapy due to their specific binding abilities.
Adjuvant is an immunological agent which enhances the body's immune response to an antigen.
Adjuvants may be added to a vaccine to boost the immune response to produce more antibodies and longer-lasting immunity, thus minimizing the dose of antigen needed to the vaccine.
Adjuvants are used in combination with a specific antigen that produced a more robust immune response than the antigen can do alone.
It includes general introduction to antibodies; Monoclonal antibodies; comparison between Polyclonal & Monoclonal antibodies; Hybridoma Technology & Hyridoma Selection; advantages & disadvantages of mABs; Applications of mABs; Recombinant Monoclonal antibodies production through Antibody Engineering.
There are 5 major antibody isotypes - IgM, IgD, IgG, IgE, and IgA - which differ based on their heavy chain. The heavy chain determines the isotype and can be mu, delta, gamma, epsilon, or alpha. Light chains can be either kappa or lambda with any isotype. IgG is the most abundant in humans while IgE is the least. Isotypes are located in the constant region of the heavy and light chains. Allotypes are specified by allelic forms of immunoglobulin genes and are also in the constant regions. Idiotypes are unique epitopes located in the variable regions of individual antibody molecules.
This document discusses monoclonal antibodies (mAbs). It provides a 3 sentence summary:
Monoclonal antibodies are identical antibodies produced by identical immune cells that are clones of a single parent cell. They are produced through the fusion of antibody-producing B cells with myeloma cells to form a hybridoma that replicates indefinitely. This process allows for the mass production of antibodies that are specific to a single antigen or epitope.
Monoclonal antibodies are produced through hybridoma technology, which involves fusing antibody-producing B-lymphocytes with myeloma cells to create a hybrid cell that can produce antibodies indefinitely. This produces a single type of antibody directed against a specific antigen. While monoclonal antibodies produced in mice can be used for in vitro applications, human monoclonal antibodies are preferred for human therapies due to reduced immunological complications. Methods to produce human monoclonal antibodies include viral transformation of human B-lymphocytes and using SCID mice or transgenic mice to generate human antibodies. Monoclonal antibodies have diagnostic, therapeutic, research, and industrial applications.
Hybridoma cells are created by fusing B lymphocytes from immunized mice with myeloma cells. This fusion produces a hybrid cell that is selected and screened to produce monoclonal antibodies against a specific antigen. The hybridoma technique allows for large-scale, indefinite production of these identical monoclonal antibodies, which have various medical uses such as in treating cancer, rheumatoid arthritis, and cardiovascular diseases. Hybridoma cells can be cultured in vitro or grown in vivo in mice to produce monoclonal antibodies.
Monoclonal antibodies (MAbs) are antibodies that are identical and bind to the same epitope. The document discusses the history and development of MAb technology, including the hybridoma technique developed by Kohler and Milstein. It describes how MAbs are produced through cell fusion, screening, and cloning. The document outlines applications of MAbs in diagnostics, such as cancer detection and imaging, and therapeutics, including cancer treatment. MAbs provide advantages over polyclonal antibodies in being highly specific and reproducible.
Adjuvant is an immunological agent which enhances the body's immune response to an antigen.
Adjuvants may be added to a vaccine to boost the immune response to produce more antibodies and longer-lasting immunity, thus minimizing the dose of antigen needed to the vaccine.
Adjuvants are used in combination with a specific antigen that produced a more robust immune response than the antigen can do alone.
It includes general introduction to antibodies; Monoclonal antibodies; comparison between Polyclonal & Monoclonal antibodies; Hybridoma Technology & Hyridoma Selection; advantages & disadvantages of mABs; Applications of mABs; Recombinant Monoclonal antibodies production through Antibody Engineering.
There are 5 major antibody isotypes - IgM, IgD, IgG, IgE, and IgA - which differ based on their heavy chain. The heavy chain determines the isotype and can be mu, delta, gamma, epsilon, or alpha. Light chains can be either kappa or lambda with any isotype. IgG is the most abundant in humans while IgE is the least. Isotypes are located in the constant region of the heavy and light chains. Allotypes are specified by allelic forms of immunoglobulin genes and are also in the constant regions. Idiotypes are unique epitopes located in the variable regions of individual antibody molecules.
This document discusses monoclonal antibodies (mAbs). It provides a 3 sentence summary:
Monoclonal antibodies are identical antibodies produced by identical immune cells that are clones of a single parent cell. They are produced through the fusion of antibody-producing B cells with myeloma cells to form a hybridoma that replicates indefinitely. This process allows for the mass production of antibodies that are specific to a single antigen or epitope.
Monoclonal antibodies are produced through hybridoma technology, which involves fusing antibody-producing B-lymphocytes with myeloma cells to create a hybrid cell that can produce antibodies indefinitely. This produces a single type of antibody directed against a specific antigen. While monoclonal antibodies produced in mice can be used for in vitro applications, human monoclonal antibodies are preferred for human therapies due to reduced immunological complications. Methods to produce human monoclonal antibodies include viral transformation of human B-lymphocytes and using SCID mice or transgenic mice to generate human antibodies. Monoclonal antibodies have diagnostic, therapeutic, research, and industrial applications.
Hybridoma cells are created by fusing B lymphocytes from immunized mice with myeloma cells. This fusion produces a hybrid cell that is selected and screened to produce monoclonal antibodies against a specific antigen. The hybridoma technique allows for large-scale, indefinite production of these identical monoclonal antibodies, which have various medical uses such as in treating cancer, rheumatoid arthritis, and cardiovascular diseases. Hybridoma cells can be cultured in vitro or grown in vivo in mice to produce monoclonal antibodies.
Monoclonal antibodies (MAbs) are antibodies that are identical and bind to the same epitope. The document discusses the history and development of MAb technology, including the hybridoma technique developed by Kohler and Milstein. It describes how MAbs are produced through cell fusion, screening, and cloning. The document outlines applications of MAbs in diagnostics, such as cancer detection and imaging, and therapeutics, including cancer treatment. MAbs provide advantages over polyclonal antibodies in being highly specific and reproducible.
1. Antigen processing and presentation involves degradation of antigens into peptides, association of peptides with MHC molecules, and display of peptide-MHC complexes on the cell surface for recognition by T cells.
2. There are two main pathways of antigen processing - exogenous antigens that enter the cell are processed through the endocytic pathway while endogenous antigens are processed through the cytosolic pathway.
3. In the cytosolic pathway, antigens are degraded by the proteasome and transported by TAP into the ER where they can bind to MHC class I molecules. In the endocytic pathway, exogenous antigens internalized into vesicles are degraded into peptides that bind MHC class II molecules.
Inbred mouse strains and adoptive transfer systems are commonly used experimental models in immunology research. Inbred strains allow researchers to study immune responses without genetic variability between subjects. The adoptive transfer technique involves irradiating mice to eliminate their immune cells, then transferring immune cells from other mice to study isolated responses. SCID mice, which lack mature immune cells, accept grafts from other species and are used to study human immune cell development through implantation of human tissues. These animal models and cell culture systems provide controlled experimental platforms for immunological research.
This document discusses different types of vaccines including synthetic peptide vaccines, recombinant antigen vaccines, and vector vaccines. Synthetic peptide vaccines use short peptide fragments to induce an immune response. Recombinant antigen vaccines produce antigens using DNA technology by inserting genes into host cells. Vector vaccines use non-pathogenic viruses or bacteria as vectors to deliver genes encoding antigens to stimulate immunity. Examples of extensively used viral vectors include vaccinia virus and adenovirus. Two vector vaccines are being developed against coronaviruses by using different viral vectors to deliver spike and nucleocapsid proteins.
Polyclonal antibodies are produced by injecting an antigen into a host animal which causes its immune system to produce various antibodies that recognize different epitopes of the antigen. The antibodies are then purified from the animal's blood plasma. Polyclonal antibodies are a heterogeneous mixture that can bind to multiple epitopes of the same antigen, whereas monoclonal antibodies are all clones that recognize the same epitope.
ANTIGEN, HAPTEN, ALL TYPES OF ANTIGENS, IMMUNOGEN , ATTRIBUTES OF ANTIGENICITY, DETERMINANTS OF ANTIGENICITY,
IMMUNOLOGY KUBY, MEDICAL MICROBIOLOGY & IMMUNOLOGY OF PANIKER , LIPPINCOTT'S IMMUNOLOGY, OTHER SOURCES.
This document summarizes antigen processing and presentation. It discusses that antigen presenting cells such as macrophages, dendritic cells, and B cells express class II MHC molecules and provide co-stimulatory signals to activate T helper cells. These cells internalize antigens through phagocytosis or endocytosis, degrade them into peptides, and present the peptides bound to class II MHC on their surface. The document also describes the major histocompatibility complex and the roles of class I and class II MHC molecules in antigen presentation to T cells. It outlines the exogenous and endogenous antigen processing pathways, how exogenous antigens are presented by class II MHC and endogenous antigens by class I MHC.
Hybridoma technology allows scientists to produce monoclonal antibodies at scale. It involves fusing antibody-producing plasma cells from immunized mice with myeloma cancer cells, creating a hybridoma cell line that is immortal and continuously produces identical monoclonal antibodies. This overcomes the short lifespan of plasma cells. The hybridoma cells are selected and isolated using HAT media, which only the hybridomas can survive in due to possessing a key enzyme. This technique has generated monoclonal antibodies useful for diagnosing and treating various diseases.
The document summarizes the process of producing monoclonal antibodies (mAbs) through hybridoma technology. It involves immunizing an animal, usually a mouse, to elicit an immune response. B cells from the animal's spleen are then fused with myeloma cells to generate immortal hybridoma cells. These hybridoma cells are screened and selected in HAT medium to identify clones that produce the desired mAb. The selected clones are then subjected to further characterization and mass production methods.
Monoclonal and polyclonal antibodies can be produced through different methods. Monoclonal antibodies are produced using hybridoma technology, which involves fusing myeloma cells with antibody-producing B cells to create immortal hybridoma cell lines. Kohler and Milstein developed this technique in 1975. Polyclonal antibodies involve immunizing an animal to produce a mixture of antibodies against various epitopes of an antigen. Monoclonal antibodies are highly specific to a single epitope, while polyclonal antibodies detect multiple epitopes but with less specificity. Monoclonal antibodies provide an unlimited supply of consistent, specific antibodies and are widely used in research and therapeutic applications.
The document summarizes Burnet's clonal selection theory of antibody production. It explains that according to this theory, lymphocyte stem cells randomly differentiate to produce mature B and T cells, each with a unique antigen receptor. When a B cell encounters the antigen it recognizes, it activates the B cell clone and causes it to proliferate and differentiate into plasma cells that secrete antibodies with the same specificity as the parental B cell receptor. The theory explains how the immune system produces antibodies and memory cells that allow for a rapid secondary immune response upon reexposure to the same antigen.
The document discusses the generation of antibody diversity in the immune system. It explains that there are millions of possible antigens but only a small number of immunoglobulin genes in our genome. Through seven mechanisms, including multiple germline genes, combinatorial V(D)J joining, junctional flexibility, and somatic hypermutation, the immune system is able to generate a diverse repertoire of antibodies against all potential antigens from a limited set of gene segments. These mechanisms operate during B cell development and maturation in the bone marrow and lymphoid tissues.
The document discusses the history and development of vaccines. It begins with early discoveries in the 18th-19th centuries relating to smallpox and rabies vaccines. It then outlines major vaccine discoveries from the 1890s-1990s for diseases such as diphtheria, polio, measles, and hepatitis B. The document also describes different types of traditional and modern vaccines, including how they are prepared and the microorganisms they contain. It provides details on live attenuated, inactivated, subunit, and viral vector vaccines.
The document summarizes the humoral immune response. It involves B cells producing antibodies that destroy extracellular microorganisms and prevent spread of intracellular infections. The process begins when a bacterium is phagocytosed by an antigen presenting cell. The antigen is processed and displayed on the cell surface. This activates helper T cells, which trigger B cell activation and antibody production. The antibodies then bind to antigens on microorganisms, marking them for destruction by immune cells and complement proteins.
Monoclonal antibodies are identical immunoglobulins generated from a single B-cell clone that recognize a unique epitope on a single antigen. They have various applications including diagnostic applications using biochemical analysis and diagnostic imaging, therapeutic applications as direct treatment agents and targeting agents, and protein purification. Monoclonal antibodies are produced by fusing B cells from an immunized animal with myeloma cells to generate hybridomas that can produce the monoclonal antibody indefinitely.
Mechanism of vd(j) recombination and generation of antibody diversityKayeen Vadakkan
The document summarizes the mechanism of V(D)J recombination and generation of antibody diversity. It discusses:
1) How V(D)J recombination involves rearrangement of one V, D (only in heavy chains), and J gene segment in B and T lymphocytes, bringing them under the control of regulatory elements.
2) The recognition signals and rearrangement process, which involves double stranded breaks and joining of coding ends.
3) The four main stages of V(D)J recombination - synapsis, cleavage, hairpin opening and end processing, and joining.
4) The seven means by which antibody diversity is generated - multiple gene segments, combinatorial joining, junctional flexibility
1. The major histocompatibility complex (MHC) helps the immune system recognize foreign substances. It is expressed on nearly all cells and plays a crucial role in organ transplant compatibility.
2. MHC molecules are classified into three types - MHC class I presents antigens to T cells within cells, MHC class II presents antigens to T cells between cells, and MHC class III encodes proteins unrelated to antigen presentation.
3. Antigens are processed through two pathways - the cytosolic pathway for endogenous antigens and the endocytic pathway for exogenous antigens - and bound to MHC molecules for presentation to T cells, which triggers an immune response against foreign or transplanted tissues that do not match the recipient's MHC.
The document summarizes the key mechanisms by which the human immune system generates a diverse repertoire of antibodies from a relatively small number of genes. It describes the somatic variation theory where mutation and recombination of immunoglobulin genes in somatic cells results in high antibody diversity. It explains processes like V(D)J recombination of light and heavy chain genes, junctional diversity, allelic exclusion, somatic hypermutation, and class switching which all contribute to antibody diversity.
Monoclonal antibodies (mAbs) have various applications including diagnostic, therapeutic, and catalytic uses. Diagnostically, mAbs are used in pregnancy tests, disease diagnostics, and immunohistochemistry. Therapeutically, mAbs treat cancer, transplant rejection, and autoimmune diseases by mechanisms like enhancing immune response, blocking growth signals, inhibiting angiogenesis or cytokines. Some FDA-approved mAbs for cancer include rituximab, trastuzumab, and bevacizumab. Conjugated mAbs can deliver toxins or radiation directly to tumors. mAbs also have applications in areas like organ transplantation, autoimmune diseases, and drug targeting.
Monoclonal antibodies are produced from single clones of B cells and recognize a specific antigen. They have advantages over polyclonal antibodies like homogeneity, specificity, and unlimited production. Monoclonal antibodies are created through cell fusion between B cells and myeloma cells to form immortal hybridoma cells that produce the same antibody. They have various medical uses including cancer therapy, diagnostics, and treatment of autoimmune disorders.
Monoclonal antibodies are identical antibodies produced by a single clone of B cells that recognize a specific epitope on an antigen. They are produced through the fusion of B cells from an immunized animal with myeloma cells to form a hybridoma. This hybridoma will continuously secrete the same monoclonal antibody. Monoclonal antibodies have various diagnostic and therapeutic applications including use in biochemical assays, diagnostic imaging, cancer treatment, and protein purification due to their high specificity for targets.
1. Antigen processing and presentation involves degradation of antigens into peptides, association of peptides with MHC molecules, and display of peptide-MHC complexes on the cell surface for recognition by T cells.
2. There are two main pathways of antigen processing - exogenous antigens that enter the cell are processed through the endocytic pathway while endogenous antigens are processed through the cytosolic pathway.
3. In the cytosolic pathway, antigens are degraded by the proteasome and transported by TAP into the ER where they can bind to MHC class I molecules. In the endocytic pathway, exogenous antigens internalized into vesicles are degraded into peptides that bind MHC class II molecules.
Inbred mouse strains and adoptive transfer systems are commonly used experimental models in immunology research. Inbred strains allow researchers to study immune responses without genetic variability between subjects. The adoptive transfer technique involves irradiating mice to eliminate their immune cells, then transferring immune cells from other mice to study isolated responses. SCID mice, which lack mature immune cells, accept grafts from other species and are used to study human immune cell development through implantation of human tissues. These animal models and cell culture systems provide controlled experimental platforms for immunological research.
This document discusses different types of vaccines including synthetic peptide vaccines, recombinant antigen vaccines, and vector vaccines. Synthetic peptide vaccines use short peptide fragments to induce an immune response. Recombinant antigen vaccines produce antigens using DNA technology by inserting genes into host cells. Vector vaccines use non-pathogenic viruses or bacteria as vectors to deliver genes encoding antigens to stimulate immunity. Examples of extensively used viral vectors include vaccinia virus and adenovirus. Two vector vaccines are being developed against coronaviruses by using different viral vectors to deliver spike and nucleocapsid proteins.
Polyclonal antibodies are produced by injecting an antigen into a host animal which causes its immune system to produce various antibodies that recognize different epitopes of the antigen. The antibodies are then purified from the animal's blood plasma. Polyclonal antibodies are a heterogeneous mixture that can bind to multiple epitopes of the same antigen, whereas monoclonal antibodies are all clones that recognize the same epitope.
ANTIGEN, HAPTEN, ALL TYPES OF ANTIGENS, IMMUNOGEN , ATTRIBUTES OF ANTIGENICITY, DETERMINANTS OF ANTIGENICITY,
IMMUNOLOGY KUBY, MEDICAL MICROBIOLOGY & IMMUNOLOGY OF PANIKER , LIPPINCOTT'S IMMUNOLOGY, OTHER SOURCES.
This document summarizes antigen processing and presentation. It discusses that antigen presenting cells such as macrophages, dendritic cells, and B cells express class II MHC molecules and provide co-stimulatory signals to activate T helper cells. These cells internalize antigens through phagocytosis or endocytosis, degrade them into peptides, and present the peptides bound to class II MHC on their surface. The document also describes the major histocompatibility complex and the roles of class I and class II MHC molecules in antigen presentation to T cells. It outlines the exogenous and endogenous antigen processing pathways, how exogenous antigens are presented by class II MHC and endogenous antigens by class I MHC.
Hybridoma technology allows scientists to produce monoclonal antibodies at scale. It involves fusing antibody-producing plasma cells from immunized mice with myeloma cancer cells, creating a hybridoma cell line that is immortal and continuously produces identical monoclonal antibodies. This overcomes the short lifespan of plasma cells. The hybridoma cells are selected and isolated using HAT media, which only the hybridomas can survive in due to possessing a key enzyme. This technique has generated monoclonal antibodies useful for diagnosing and treating various diseases.
The document summarizes the process of producing monoclonal antibodies (mAbs) through hybridoma technology. It involves immunizing an animal, usually a mouse, to elicit an immune response. B cells from the animal's spleen are then fused with myeloma cells to generate immortal hybridoma cells. These hybridoma cells are screened and selected in HAT medium to identify clones that produce the desired mAb. The selected clones are then subjected to further characterization and mass production methods.
Monoclonal and polyclonal antibodies can be produced through different methods. Monoclonal antibodies are produced using hybridoma technology, which involves fusing myeloma cells with antibody-producing B cells to create immortal hybridoma cell lines. Kohler and Milstein developed this technique in 1975. Polyclonal antibodies involve immunizing an animal to produce a mixture of antibodies against various epitopes of an antigen. Monoclonal antibodies are highly specific to a single epitope, while polyclonal antibodies detect multiple epitopes but with less specificity. Monoclonal antibodies provide an unlimited supply of consistent, specific antibodies and are widely used in research and therapeutic applications.
The document summarizes Burnet's clonal selection theory of antibody production. It explains that according to this theory, lymphocyte stem cells randomly differentiate to produce mature B and T cells, each with a unique antigen receptor. When a B cell encounters the antigen it recognizes, it activates the B cell clone and causes it to proliferate and differentiate into plasma cells that secrete antibodies with the same specificity as the parental B cell receptor. The theory explains how the immune system produces antibodies and memory cells that allow for a rapid secondary immune response upon reexposure to the same antigen.
The document discusses the generation of antibody diversity in the immune system. It explains that there are millions of possible antigens but only a small number of immunoglobulin genes in our genome. Through seven mechanisms, including multiple germline genes, combinatorial V(D)J joining, junctional flexibility, and somatic hypermutation, the immune system is able to generate a diverse repertoire of antibodies against all potential antigens from a limited set of gene segments. These mechanisms operate during B cell development and maturation in the bone marrow and lymphoid tissues.
The document discusses the history and development of vaccines. It begins with early discoveries in the 18th-19th centuries relating to smallpox and rabies vaccines. It then outlines major vaccine discoveries from the 1890s-1990s for diseases such as diphtheria, polio, measles, and hepatitis B. The document also describes different types of traditional and modern vaccines, including how they are prepared and the microorganisms they contain. It provides details on live attenuated, inactivated, subunit, and viral vector vaccines.
The document summarizes the humoral immune response. It involves B cells producing antibodies that destroy extracellular microorganisms and prevent spread of intracellular infections. The process begins when a bacterium is phagocytosed by an antigen presenting cell. The antigen is processed and displayed on the cell surface. This activates helper T cells, which trigger B cell activation and antibody production. The antibodies then bind to antigens on microorganisms, marking them for destruction by immune cells and complement proteins.
Monoclonal antibodies are identical immunoglobulins generated from a single B-cell clone that recognize a unique epitope on a single antigen. They have various applications including diagnostic applications using biochemical analysis and diagnostic imaging, therapeutic applications as direct treatment agents and targeting agents, and protein purification. Monoclonal antibodies are produced by fusing B cells from an immunized animal with myeloma cells to generate hybridomas that can produce the monoclonal antibody indefinitely.
Mechanism of vd(j) recombination and generation of antibody diversityKayeen Vadakkan
The document summarizes the mechanism of V(D)J recombination and generation of antibody diversity. It discusses:
1) How V(D)J recombination involves rearrangement of one V, D (only in heavy chains), and J gene segment in B and T lymphocytes, bringing them under the control of regulatory elements.
2) The recognition signals and rearrangement process, which involves double stranded breaks and joining of coding ends.
3) The four main stages of V(D)J recombination - synapsis, cleavage, hairpin opening and end processing, and joining.
4) The seven means by which antibody diversity is generated - multiple gene segments, combinatorial joining, junctional flexibility
1. The major histocompatibility complex (MHC) helps the immune system recognize foreign substances. It is expressed on nearly all cells and plays a crucial role in organ transplant compatibility.
2. MHC molecules are classified into three types - MHC class I presents antigens to T cells within cells, MHC class II presents antigens to T cells between cells, and MHC class III encodes proteins unrelated to antigen presentation.
3. Antigens are processed through two pathways - the cytosolic pathway for endogenous antigens and the endocytic pathway for exogenous antigens - and bound to MHC molecules for presentation to T cells, which triggers an immune response against foreign or transplanted tissues that do not match the recipient's MHC.
The document summarizes the key mechanisms by which the human immune system generates a diverse repertoire of antibodies from a relatively small number of genes. It describes the somatic variation theory where mutation and recombination of immunoglobulin genes in somatic cells results in high antibody diversity. It explains processes like V(D)J recombination of light and heavy chain genes, junctional diversity, allelic exclusion, somatic hypermutation, and class switching which all contribute to antibody diversity.
Monoclonal antibodies (mAbs) have various applications including diagnostic, therapeutic, and catalytic uses. Diagnostically, mAbs are used in pregnancy tests, disease diagnostics, and immunohistochemistry. Therapeutically, mAbs treat cancer, transplant rejection, and autoimmune diseases by mechanisms like enhancing immune response, blocking growth signals, inhibiting angiogenesis or cytokines. Some FDA-approved mAbs for cancer include rituximab, trastuzumab, and bevacizumab. Conjugated mAbs can deliver toxins or radiation directly to tumors. mAbs also have applications in areas like organ transplantation, autoimmune diseases, and drug targeting.
Monoclonal antibodies are produced from single clones of B cells and recognize a specific antigen. They have advantages over polyclonal antibodies like homogeneity, specificity, and unlimited production. Monoclonal antibodies are created through cell fusion between B cells and myeloma cells to form immortal hybridoma cells that produce the same antibody. They have various medical uses including cancer therapy, diagnostics, and treatment of autoimmune disorders.
Monoclonal antibodies are identical antibodies produced by a single clone of B cells that recognize a specific epitope on an antigen. They are produced through the fusion of B cells from an immunized animal with myeloma cells to form a hybridoma. This hybridoma will continuously secrete the same monoclonal antibody. Monoclonal antibodies have various diagnostic and therapeutic applications including use in biochemical assays, diagnostic imaging, cancer treatment, and protein purification due to their high specificity for targets.
This document provides an overview of monoclonal antibodies and gene therapy. It discusses the discovery of monoclonal antibodies by Kohler and Milstein in 1975. It also describes the multi-step process of producing monoclonal antibodies through cell fusion and hybridoma technology. Several types of monoclonal antibodies are outlined, along with their purification techniques and therapeutic applications in cancer treatment and other diseases. Gene therapy approaches including ex vivo and in vivo methods are briefly introduced.
This document discusses monoclonal antibodies, including their discovery and types. It provides details on:
- George Kohler and Cesar Milstein discovered hybridoma technology in 1975, which enables the production of monoclonal antibodies from a single clone of B cells.
- There are four main types of monoclonal antibodies based on their origin: murine, chimeric, humanized, and fully human. Hybridoma technology fuses B cells with myeloma cells to produce monoclonal antibodies indefinitely in culture.
- Monoclonal antibodies can be "naked" or conjugated to drugs, toxins, or radioactive particles to target cancers. They work by binding to specific antigens on cells and triggering immune responses or delivering cytotoxic payloads to targeted
This document summarizes monoclonal antibodies and gene therapy. It discusses the discovery of monoclonal antibodies by Kohler and Milstein in 1975. It describes the production process of monoclonal antibodies which involves immunizing mice, fusing spleen cells with myeloma cells to form hybridomas, and cloning cell lines. The document also discusses types of monoclonal antibodies including murine, chimeric, and humanized, as well as applications in cancer therapy, diagnostics, and immunosuppression. Gene therapy techniques like ex vivo and in vivo delivery are summarized along with strategies for cancer like suicide gene therapy using thymidine kinase.
Production and applications of monoclonal antibodiesKaayathri Devi
production and applications of monoclonal antibodies, monoclonal antibodies ,applications of monoclonal antibodies, production of monoclonal antibodies,
Monoclonal antibodies in cancer treatment By Ankit TribhuvaneMumbai University
Monoclonal antibodies are identical antibodies produced by a single clone of B cells that specifically bind to target cells. They can be used for cancer therapy by triggering immune system attacks on cancer cells, blocking growth signals, or preventing new blood vessel formation. Monoclonal antibodies are produced through hybridoma technology, fusing B cells with myeloma cells. This produces immortal clones that secrete identical antibodies. Monoclonal antibodies have applications in cancer diagnosis and treatment, with therapeutic antibodies targeting tumors through mechanisms like antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity.
Monoclonal Antibody-Preparation & Application - MPH201T.pptxRAHUL PAL
Monoclonal antibodies (mAbs) are proteins produced by a single type of B cell. They are identical to each other and recognize a specific antigen. Antigens are molecules that the body's immune system recognizes as foreign. When an antigen binds to a monoclonal antibody, it triggers a series of reactions that can lead to the destruction of the antigen.
Monoclonal antibodies can be used to treat a variety of diseases, including cancer, autoimmune diseases, and infections. They are also used in research and diagnostics.
Monoclonal antibodies (mAbs) are identical antibodies produced by a single clone of immune cells that are all clones of the same parent cell. mAbs can be produced against almost any substance and are important tools in biochemistry and medicine. They are produced through the fusion of antibody-producing cells with myeloma cells to form hybridomas that produce identical antibodies. mAbs have applications in research, diagnostics, and therapy due to their specificity and ability to detect or purify target substances.
Monoclonal antibodies are produced from single clones of B cells and bind to a specific epitope. Georges Köhler and Cesar Milstein developed hybridoma technology in 1975, which involves fusing B cells with myeloma cells to generate immortal hybridoma cell lines that secrete monoclonal antibodies. Hybridomas are selected by growing the cells in HAT medium, which kills unfused cells but allows hybridomas to survive and proliferate indefinitely. Monoclonal antibodies have numerous applications including cancer immunotherapy, diagnostic tests, and treatment of various diseases due to their high specificity and stability.
Monoclonal antibodies are identical antibodies produced by a single B cell clone that recognize a specific epitope. They are produced through the fusion of B cells from an immunized animal with myeloma cells to form a hybridoma cell line. Monoclonal antibodies have various applications, including use in diagnostic tests to detect substances like hormones and tumor markers, diagnostic imaging by delivering radioisotopes to target areas, and directly targeting diseases or purifying proteins through immunoaffinity chromatography. Their specificity and ability to target single epitopes makes them useful research and medical tools.
This document discusses monoclonal antibodies, including their structure, types, methods of preparation, applications, and marketed products. Monoclonal antibodies are homogeneous antibodies produced by identical immune cells that are clones of a single parent cell. They are prepared using the hybridoma technology which involves immunizing an animal, fusing immune cells with myeloma cells to form hybridomas, screening and selecting hybridomas that produce the desired monoclonal antibody. Monoclonal antibodies have various therapeutic applications in areas such as cancer, autoimmune diseases, organ transplants, and infections. They are also used for diagnostic purposes.
Monoclonal antibodies & hybridoma technologyAjay Dominic
Monoclonal antibodies are produced from a single clone of cells and bind to the same epitope. They are produced using hybridoma technology which involves fusing antibody-secreting B cells with myeloma cells to form immortal hybridoma cells that secrete monoclonal antibodies. This technique was developed by Kohler and Milstein in 1975 and they received the Nobel Prize.
Monoclonal antibodies are produced through the hybridoma technology developed by Kohler and Milstein in 1975. This involves fusing B cells from an immunized animal with myeloma cells to produce hybridomas that secrete a single antibody specific to the target antigen. The antibodies can be screened and a clone selected to mass produce monoclonal antibodies that recognize a single epitope. Monoclonal antibodies have various applications including disease diagnosis, immunotherapy, and immunosuppression for organ transplants. While powerful tools, limitations include potential immunogenicity and high production costs.
Production of monoclonal antibodies and applications in therapy and diagnosisAhmed Madni
Monoclonal antibodies are identical antibodies produced by a single clone of cells that bind to a specific epitope. They are produced through the fusion of antibody-producing B cells with myeloma cells, generating hybridoma cells that can produce antibodies indefinitely. Monoclonal antibodies have applications in therapy and diagnosis, including detecting antigens through techniques like ELISA, purifying proteins, and treating cancers by delivering toxins or radioisotopes to tumor cells. Advances in engineering have reduced issues like human anti-mouse antibody responses by creating chimeric or humanized antibodies.
What are Antibody
Monoclonal Antibody (mAb)
Structure of mAb
Types of Monoclonal Antibody (mAb)
Preparation of Monoclonal Antibody
Hybridoma Technique, Phage display Technique
Application of Monoclonal Antibody
Advantage and Disadvantage of Monoclonal Antibody
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2. INTRODUCTION
MONOCLONAL ANTIBODY
Antibodies which are derived from a single clone of plasma cells, all
antibodies having the same antigen specificity, i.e. produced against a single
epitope of an antigen is called monoclonal antibody
POLYCONAL ANTIBODY
Antibodies which are derived from a multiple clone of plasma cells, all
antibodies having the different antigen specificity, i.e. produced against a
various epitope of an antigen is called polyclonal antibody
3. Monoclonal antibodies have revolutionized the field of medicine, as they
can be designed to target specific cells or proteins in the body. This
specificity makes them incredibly useful for diagnosing and treating
diseases such as cancer, autoimmune disorders, and infectious diseases.
In addition to their specificity, monoclonal antibodies also have low
toxicity, making them ideal for use in humans. They can be used to deliver
drugs directly to cancer cells they are also called as Magic bullets.
4. HOW ARE MONOCLONALANTIBODIES PRODUCED?
• Monoclonal antibodies are produced by hybridoma technology.
• This technology was discovered in 1975 by George Kohler of west
Germany and Cesal Milstein of Argentina (Nobel prize, 1984).
Kohler experiment
• Kohler (1974) successfully produced hybridoma by fusing a P3
myeloma cell and a lymphocyte (from the spleen of a mouse)
immunized against the sheep red blood cells.
• These hybridomas retained the property of immortality of the
myeloma cells as well as that of secreting an antibody specific for
an sheep red blood cells antigen.
5. MONOCLONALANTIBODY PRODUCTION PROCESS
1. Purity form of immunogen
2. Choice of animals
3. Immunization
4. Separation of lymphocytes
5. Selection of myeloma cell line
6. Immortalization by fusion with tumor cells
7. Selection of hybridoma cells
8. Isolation of a monoclonal antibody producing hybridoma cell
9. Purification of desired antibodies
6. Purity and Form of Immunogen
The purity of the antigen to be used as immunogen is crucial for generation
of monoclonal antibodies.
Molecules of low molecular weight (1000 Daltons) are poor immunogens
and have to be coupled to larger immunogenic molecules.
Aggregated and particulate antigens elicit stronger responses.
Adjuvants help to achieve stronger responses. The most common carriers are
albumins, keyhole limpet hemocyanin, fowl gamma globulin and
synthetic polypeptides.
Selection of Myeloma Cell Line
The myeloma cell line used must itself not be capable of synthesizing
antibody otherwise hybridoma cell line will produce a mixture of antibodies.
HPRT-negative myeloma cell line should be selected
7. Immunization procedures
Generally the antigen is injected subcutaneously or into the peritoneal cavity of
the animal along with an adjuvant to stimulate the immune system.
Sacrifice of animal and separation of lymphocytes
Three days after the final dose of antigen has been given intravenously to
immunize the animal, the latter is killed.
The spleen of the killed animal is removed aseptically and gently disrupted to
release the spleen fluid containing lymphocytes and red blood cells.
The lymphocytes are separated from the spleen fluid (and red blood cells) by
density gradient centrifugation, and washed.
9. Immortalization by Fusion with Tumor Cells
The cell fusion process between myeloma cells and spleen lymphocytes is
usually induced in the presence of polyethylene glycol (PEG-1500)
The first fusion experiments were performed with Sendai virus as a
fusogenic agent.
Both regents aggregate the cells which eventually lead to fusion of cells.
Selection of Hybridoma Cells
The basis of hybridoma technology was the development of suitable
myeloma mutant cell lines that are non-antibody secreting and deficient in
the enzyme Hypoxanthine Guanine Phosphoribosyl Transferase
(HGPRT) and are not able to grow in toxic tissue culture.
The enzyme HGPRT is essential for DNA synthesis (by purine salvage
pathway) after fusion of these myeloma cells with lymphocytes.
11. The HAT (Hypoxanthine-Aminopterin-Thymidine) Medium is used for metabolic
selection of fused cells.
The medium containing HAT will eliminate the unfused myeloma cells and allow the
growth of hybridomas which harbour the gene(s) for HGRPT from the parent lymphocytes.
Principle of HAT Medium
The HGPRT myeloma cells die off as aminopterin blocks the main pathway of DNA
synthesis, i.e., de novo pathway of DNA synthesis by inhibiting the activity of
dihydrofolate reductase. In hybrid cells, spleen cells contribute the functional HGPRT
enzyme necessary to overcome the aminopterin block.
Unfused Lymphocyte cells are eliminated due to poor growth in vitro.
12. Isolation of a Monoclonal antibody Producing Hybridoma cell
If all the hybridoma cells that have been selected using HAT-medium, a polyclonal antibody mixture
would be obtained.
Consequently, a single antibody (monoclonal antibody) producing hybridoma cells need to be isolated
and grown individually.
This is done by diluting a suspension of hybridoma cells to such an extent that individual aliquots
contain, on an average, only one cell.
Such cells are transferred to separate fresh media for growth.
Each mass of hybridoma cells (clone) produced from a single parent hybridoma cell is now examined
to determine whether it produces the desired monoclonal antibody thought the affinity
chromatography or precipitation techniques.
13. Isolation of a
monoclonal antibody
producing hybridoma
cell
Selection of
hybridoma cells
Immortalizati
on by fusion
with tumor
cells
Sacrifice of
animal and
separation of
lymphocytes
Immunization
MONOCLONAL
ANTIBODY
PRODUCTION
PROCESS
14. APPLICATION
Therapeutic Applications. Monoclonal antibodies are used as therapeutic agents
to treat a wide range of diseases, including: Cancer Treatment, Autoimmune
Disorders, Infectious Diseases and Transplant Medicine.
Diagnosis and Monitoring. Monoclonal antibodies are used in diagnostic tests to
identify specific antigens or markers associated with diseases. For instance,
pregnancy tests use mAbs to detect human chorionic gonadotropin (hCG) in urine.
Research Tools. mAbs are essential tools in biomedical research. They help
scientists study various proteins, receptors, and cells by specifically targeting them.
This aids in understanding their functions, interactions, and roles in diseases
15. Immunotherapy. Apart from cancer treatment, mAbs are used in immunotherapy to stimulate or enhance
the body's immune response against diseases. They can activate immune cells, like T cells, to target
infected or cancerous cells more effectively.
Drug Delivery. mAbs can serve as carriers for drugs or imaging agents, specifically delivering them to
targeted cells or tissues, reducing side effects, and improving treatment efficacy.
Imaging and Diagnostics: Monoclonal antibodies can be labeled with radioactive isotopes or other
imaging agents to detect specific cells or tissues in imaging techniques such as positron emission
tomography (PET) and single-photon emission computed tomography (SPECT).
Biotechnology and Industrial Applications. mAbs are utilized in various biotechnological processes,
including protein purification, as well as in the development of biosensors and diagnostic devices.
Passive Immunization: In certain situations, mAbs can be administered passively to provide immediate,
short-term protection against infections or toxins. This approach is used in cases of exposure to certain
diseases or as a preventive measure.
16. POLYCLONALANTIBODIES
Antibodies which are derived from a multiple clone of plasma cells, all
antibodies having the different antigen specificity, i.e. produced against a
various epitope of an antigen is called polyclonal antibody
Advantages of polyclonal antibodies.
Easy, cheap and quick preparation.
PAbs are heterogeneous, bind to a wide range of antigen epitopes.
PAbs can be made in large quantities.
17. THE GENERAL PROCEDURE TO PRODUCE POLYCLONALANTIBODIES
Selection of Purity form of immunogen
Animal selection
Immunization
Isolation of Polyclonal antibodies
18. Procedure To Produce Polyclonal Antibodies
A host Sp. is chosen for the production
of Polyclonal antibodies considering
three main aspects:-
1. Quantity of antibodies required.
2. Antigen source.
3. Final application of the
polyclonal antibodies.
1. Molecular Weight Of
Immunogen Should
Be 1000 kDa. Or
2. Have To Be Coupled
To Larger
Immunogenic
Molecules.
3. Adjuvants help to
achieve stronger
responses.
Antibodies purification
Affinity chromatography is used for
Polyclonal antibodies purification
There are two processes
Antibody / Immunoglobulin
specific.
Antigen specific purification.
Validation of
antibodies/ Quality
control.
1. concentration is
measured taking
absorbance at
280mm.
2. Purity is checked
by SDS-PAGE
3. Titer is
estimated by main
ly ELISA
20. APPLICATION
Immunoassays. Polyclonal antibodies are widely used in various immunoassays, such as enzyme-
linked immunosorbent assays (ELISA), Western blotting, and immunohistochemistry. They can be
used to detect and quantify the presence of specific antigens in biological samples.
Diagnostic testing. Polyclonal antibodies are employed in clinical diagnostic tests for detecting
diseases and infections. For example, they are used in rapid diagnostic tests for detecting viral or
bacterial infections like HIV, hepatitis, and influenza.
Therapeutic Applications. Some polyclonal antibodies have therapeutic uses. They can be
administered to patients to boost the immune response against certain pathogens or toxins.
Additionally, polyclonal antibodies may be used to neutralize toxins or pathogens in cases where
specific monoclonal antibodies are not available.
21. In Vivo Research. Polyclonal antibodies are utilized in animal studies and
research to investigate specific biological processes, identify protein
expression patterns, and examine cellular localization of proteins.
Flow Cytometry. Polyclonal antibodies labeled with fluorescent tags are
employed in flow cytometry, allowing researchers to analyze and sort cells
based on specific surface markers and intracellular antigens.
Immunoprecipitation. Polyclonal antibodies can be used to isolate and pull
down specific proteins or protein complexes from a mixture of proteins using
immunoprecipitation techniques.
22. Chromatin Immunoprecipitation (ChIP). In ChIP assays, polyclonal
antibodies are used to investigate the interactions between proteins and DNA
by isolating specific DNA sequences bound to a particular protein of interest.
Neutralization Assays. Polyclonal antibodies can be employed in
neutralization assays to determine their ability to block the activity of
pathogens, toxins, or other antigens.
Purification of Proteins. Polyclonal antibodies can be used to purify proteins
of interest through affinity chromatography, where the target protein is
specifically captured by the antibody.