What are monoclonal antibodies? advantages and disadvantages, their experiments, production of hybridoma lines, their action in Escherichia coli, and applications.
This document discusses monoclonal antibodies, including their preparation and applications. It begins with an introduction to antibodies and defines monoclonal antibodies as artificial antibodies produced from a single clone. The document then summarizes the key steps in producing monoclonal antibodies, which involves immunizing an animal, fusing B lymphocytes with myeloma cells to create immortal hybridomas, selecting antibody-producing hybridomas, culturing the cells, and extracting and purifying the monoclonal antibodies. The applications section notes that monoclonal antibodies are used for disease diagnosis, passive immunization, detecting and purifying biomolecules, and some have been authorized to treat COVID-19.
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.
This document discusses monoclonal antibody production and applications. It begins by defining monoclonal and polyclonal antibodies, noting that monoclonal antibodies are identical because they are derived from a single parent cell. It then covers the history of monoclonal antibody development, the monoclonal antibody production method involving mouse immunization and cell fusion, and the types and uses of monoclonal antibodies including diagnostic and cancer treatment applications. Potential side effects of monoclonal antibody therapy are also mentioned.
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
Monoclonal antibodies are identical antibodies produced by one type of immune cell that are clones of a single parent cell. They are produced using hybridoma technology which involves fusing antibody producing B cells from an immunized animal with myeloma tumor cells to create a hybridoma cell line. This hybridoma cell line is capable of indefinite division in culture while producing the same monoclonal antibody. The monoclonal antibodies are then purified from the culture supernatant and have various diagnostic and therapeutic applications such as cancer treatment.
Monoclonal antibodies are identical antibodies produced by a single clone of B cells that recognize the same epitope on an antigen. They can be produced in a laboratory by fusing B cells that produce a desired antibody with myeloma cells to form hybridomas. Monoclonal antibodies have many diagnostic applications as they can be used to detect specific substances like proteins, pathogens, and tumor markers. They allow for rapid diagnosis of diseases like hepatitis, influenza, and cancer. Monoclonal antibodies are also used in pregnancy tests and monitoring drug levels in the body.
Hybridoma technology for production of monoclonal antibodies.pdfsoniaangeline
The document discusses hybridoma technology for producing monoclonal antibodies. It describes the process which involves fusing antibody-producing B cells from immunized mice with myeloma tumor cells, using a chemical or electrical method. The resulting hybridoma cells are selected and cultured, retaining the antibody-producing ability of B cells and indefinite lifespan of tumor cells. The monoclonal antibodies produced can be extracted and purified for therapeutic and research applications.
PPT ON MONOCLONAL ANTIBODIES.saurabh punia.ppt.pptxSAURABH PUNIA
This document discusses monoclonal antibodies, including their production, structure, and applications. Monoclonal antibodies are produced through hybridoma technology, which involves fusing antibody-producing B cells with myeloma cells to create immortal cell lines. They have identical binding sites since they are clones of a single parent cell. The production process involves immunizing an animal, harvesting spleen cells, fusing them with myeloma cells, selecting hybridomas that produce the desired antibody, and propagating these cells to produce large amounts of monoclonal antibodies. Monoclonal antibodies have various medical applications, such as targeted delivery of cancer drugs and treatment of diseases.
This document discusses monoclonal antibodies, including their preparation and applications. It begins with an introduction to antibodies and defines monoclonal antibodies as artificial antibodies produced from a single clone. The document then summarizes the key steps in producing monoclonal antibodies, which involves immunizing an animal, fusing B lymphocytes with myeloma cells to create immortal hybridomas, selecting antibody-producing hybridomas, culturing the cells, and extracting and purifying the monoclonal antibodies. The applications section notes that monoclonal antibodies are used for disease diagnosis, passive immunization, detecting and purifying biomolecules, and some have been authorized to treat COVID-19.
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.
This document discusses monoclonal antibody production and applications. It begins by defining monoclonal and polyclonal antibodies, noting that monoclonal antibodies are identical because they are derived from a single parent cell. It then covers the history of monoclonal antibody development, the monoclonal antibody production method involving mouse immunization and cell fusion, and the types and uses of monoclonal antibodies including diagnostic and cancer treatment applications. Potential side effects of monoclonal antibody therapy are also mentioned.
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
Monoclonal antibodies are identical antibodies produced by one type of immune cell that are clones of a single parent cell. They are produced using hybridoma technology which involves fusing antibody producing B cells from an immunized animal with myeloma tumor cells to create a hybridoma cell line. This hybridoma cell line is capable of indefinite division in culture while producing the same monoclonal antibody. The monoclonal antibodies are then purified from the culture supernatant and have various diagnostic and therapeutic applications such as cancer treatment.
Monoclonal antibodies are identical antibodies produced by a single clone of B cells that recognize the same epitope on an antigen. They can be produced in a laboratory by fusing B cells that produce a desired antibody with myeloma cells to form hybridomas. Monoclonal antibodies have many diagnostic applications as they can be used to detect specific substances like proteins, pathogens, and tumor markers. They allow for rapid diagnosis of diseases like hepatitis, influenza, and cancer. Monoclonal antibodies are also used in pregnancy tests and monitoring drug levels in the body.
Hybridoma technology for production of monoclonal antibodies.pdfsoniaangeline
The document discusses hybridoma technology for producing monoclonal antibodies. It describes the process which involves fusing antibody-producing B cells from immunized mice with myeloma tumor cells, using a chemical or electrical method. The resulting hybridoma cells are selected and cultured, retaining the antibody-producing ability of B cells and indefinite lifespan of tumor cells. The monoclonal antibodies produced can be extracted and purified for therapeutic and research applications.
PPT ON MONOCLONAL ANTIBODIES.saurabh punia.ppt.pptxSAURABH PUNIA
This document discusses monoclonal antibodies, including their production, structure, and applications. Monoclonal antibodies are produced through hybridoma technology, which involves fusing antibody-producing B cells with myeloma cells to create immortal cell lines. They have identical binding sites since they are clones of a single parent cell. The production process involves immunizing an animal, harvesting spleen cells, fusing them with myeloma cells, selecting hybridomas that produce the desired antibody, and propagating these cells to produce large amounts of monoclonal antibodies. Monoclonal antibodies have various medical applications, such as targeted delivery of cancer drugs and treatment of diseases.
This document provides an overview of monoclonal antibodies including their history, development, preparation, and applications. It discusses how monoclonal antibodies are produced through the fusion of B cells and myeloma cells to form hybridomas. The document outlines the key applications of monoclonal antibodies in diagnosis (e.g. biochemical analysis, imaging), therapy (e.g. cancer, transplantation), and protein purification. It also provides examples of commercially available monoclonal antibodies for various diseases and conditions.
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 or hybridomas. They can be used for diagnostic tests and therapy. In 1975, Köhler and Milstein developed the technique of fusing myeloma cells with spleen cells from immunized mice to generate hybridomas that produce monoclonal antibodies. This provided an unlimited supply of identical antibodies against specific antigens. Monoclonal antibodies have various applications such as diagnostic tests, purification of substances, and cancer treatment when conjugated to toxins or radioisotopes. However, they can cause side effects like allergic reactions, vomiting, and diarrhea when used intravenously.
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.
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 identical immunoglobulins generated from a single B-cell clone that recognize a unique epitope on a single antigen. They are produced through the fusion of B-cells from an immunized animal with myeloma cells. This process generates hybridoma cells that can produce monoclonal antibodies indefinitely in culture. Monoclonal antibodies have various applications, including use in diagnostic tests, as therapeutic agents, and for protein purification due to their high specificity.
This document summarizes and compares the processes for producing monoclonal and polyclonal antibodies. Monoclonal antibodies are produced from a single clone of B cells and recognize a single epitope, while polyclonal antibodies are a heterogeneous mixture produced from different B cell clones that recognize multiple epitopes of the same antigen. The production of monoclonal antibodies involves immunizing an animal, fusing B cells with myeloma cells to create hybridomas, and screening for antibodies. Polyclonal antibodies are harvested directly from immunized animal serum. The document outlines the key similarities and differences between these two antibody types.
Monoclonal antibody production by hybridoma technologyHasnat Tariq
Monoclonal antibodies are identical antibodies produced by a single clone of B cells that recognize a specific epitope. They are produced through the hybridoma technology which involves fusing antibody producing B cells with myeloma cells to form a hybridoma cell line. This allows for the mass production of antibodies that recognize a single epitope. Monoclonal antibodies have various applications including use in biochemical analysis through techniques like ELISA and RIA. They can also be used for diagnostic imaging by labeling them with radioisotopes.
Monoclonal antibodies are identical antibodies produced by a single clone of B cells that are specific to a single epitope. They offer reproducible and predictable immune responses. Polyclonal antibodies are produced by multiple B cell clones and bind to multiple epitopes. Monoclonal antibodies are produced using hybridoma technology which fuses myeloma cells with antibody producing spleen cells to generate immortal hybridoma cell lines. They have various applications in diagnostics and therapeutics including cancer treatment by mechanisms like blocking receptor signaling, immunomodulation and antibody-dependent cytotoxicity.
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.
The document discusses monoclonal antibodies, including their history, preparation, evaluation, and applications. It begins with basic concepts of antibodies and the difference between monoclonal and polyclonal antibodies. The key points are:
- Monoclonal antibodies are identical antibodies produced by a single clone of B cells that target a specific epitope, whereas polyclonal antibodies target multiple epitopes and are produced by different B cell lines.
- Preparation involves immunizing an animal, fusing spleen cells containing antibody-producing B cells with myeloma cells, screening clones to identify those producing the desired antibody, and growing the selected clone indefinitely to harvest the monoclonal antibodies.
- Evaluation includes characterization of physicochemical properties, immunological properties like affinity and specificity
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
Project Report on Monoclonal antibodies By VanshikaVanshikaBeniwal
HYBRIDOMA TECHNOLOGY
Monoclonal antibodies (MAbs) are a kind of immunological instrument that has been employed in immunology, biotechnology, biochemistry, and applied biology for a protracted time.
1. Isolate B cells from recovered COVID-19 patients that produce antibodies against the virus.
2. Fuse the B cells with myeloma cells to generate immortalized hybridoma cell lines.
3. Select a hybridoma that produces the desired anti-COVID-19 antibody and sequence the antibody genes.
4. Insert the antibody genes into a mammalian expression vector and transfect CHO cells for recombinant antibody production.
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.
How to Make a Field Mandatory in Odoo 17Celine George
In Odoo, making a field required can be done through both Python code and XML views. When you set the required attribute to True in Python code, it makes the field required across all views where it's used. Conversely, when you set the required attribute in XML views, it makes the field required only in the context of that particular view.
This document provides an overview of monoclonal antibodies including their history, development, preparation, and applications. It discusses how monoclonal antibodies are produced through the fusion of B cells and myeloma cells to form hybridomas. The document outlines the key applications of monoclonal antibodies in diagnosis (e.g. biochemical analysis, imaging), therapy (e.g. cancer, transplantation), and protein purification. It also provides examples of commercially available monoclonal antibodies for various diseases and conditions.
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 or hybridomas. They can be used for diagnostic tests and therapy. In 1975, Köhler and Milstein developed the technique of fusing myeloma cells with spleen cells from immunized mice to generate hybridomas that produce monoclonal antibodies. This provided an unlimited supply of identical antibodies against specific antigens. Monoclonal antibodies have various applications such as diagnostic tests, purification of substances, and cancer treatment when conjugated to toxins or radioisotopes. However, they can cause side effects like allergic reactions, vomiting, and diarrhea when used intravenously.
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.
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 identical immunoglobulins generated from a single B-cell clone that recognize a unique epitope on a single antigen. They are produced through the fusion of B-cells from an immunized animal with myeloma cells. This process generates hybridoma cells that can produce monoclonal antibodies indefinitely in culture. Monoclonal antibodies have various applications, including use in diagnostic tests, as therapeutic agents, and for protein purification due to their high specificity.
This document summarizes and compares the processes for producing monoclonal and polyclonal antibodies. Monoclonal antibodies are produced from a single clone of B cells and recognize a single epitope, while polyclonal antibodies are a heterogeneous mixture produced from different B cell clones that recognize multiple epitopes of the same antigen. The production of monoclonal antibodies involves immunizing an animal, fusing B cells with myeloma cells to create hybridomas, and screening for antibodies. Polyclonal antibodies are harvested directly from immunized animal serum. The document outlines the key similarities and differences between these two antibody types.
Monoclonal antibody production by hybridoma technologyHasnat Tariq
Monoclonal antibodies are identical antibodies produced by a single clone of B cells that recognize a specific epitope. They are produced through the hybridoma technology which involves fusing antibody producing B cells with myeloma cells to form a hybridoma cell line. This allows for the mass production of antibodies that recognize a single epitope. Monoclonal antibodies have various applications including use in biochemical analysis through techniques like ELISA and RIA. They can also be used for diagnostic imaging by labeling them with radioisotopes.
Monoclonal antibodies are identical antibodies produced by a single clone of B cells that are specific to a single epitope. They offer reproducible and predictable immune responses. Polyclonal antibodies are produced by multiple B cell clones and bind to multiple epitopes. Monoclonal antibodies are produced using hybridoma technology which fuses myeloma cells with antibody producing spleen cells to generate immortal hybridoma cell lines. They have various applications in diagnostics and therapeutics including cancer treatment by mechanisms like blocking receptor signaling, immunomodulation and antibody-dependent cytotoxicity.
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.
The document discusses monoclonal antibodies, including their history, preparation, evaluation, and applications. It begins with basic concepts of antibodies and the difference between monoclonal and polyclonal antibodies. The key points are:
- Monoclonal antibodies are identical antibodies produced by a single clone of B cells that target a specific epitope, whereas polyclonal antibodies target multiple epitopes and are produced by different B cell lines.
- Preparation involves immunizing an animal, fusing spleen cells containing antibody-producing B cells with myeloma cells, screening clones to identify those producing the desired antibody, and growing the selected clone indefinitely to harvest the monoclonal antibodies.
- Evaluation includes characterization of physicochemical properties, immunological properties like affinity and specificity
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
Project Report on Monoclonal antibodies By VanshikaVanshikaBeniwal
HYBRIDOMA TECHNOLOGY
Monoclonal antibodies (MAbs) are a kind of immunological instrument that has been employed in immunology, biotechnology, biochemistry, and applied biology for a protracted time.
1. Isolate B cells from recovered COVID-19 patients that produce antibodies against the virus.
2. Fuse the B cells with myeloma cells to generate immortalized hybridoma cell lines.
3. Select a hybridoma that produces the desired anti-COVID-19 antibody and sequence the antibody genes.
4. Insert the antibody genes into a mammalian expression vector and transfect CHO cells for recombinant antibody production.
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.
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In Odoo, making a field required can be done through both Python code and XML views. When you set the required attribute to True in Python code, it makes the field required across all views where it's used. Conversely, when you set the required attribute in XML views, it makes the field required only in the context of that particular view.
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2. CONTENTS
• Basic concept and introduction.
• Advantages and disadvantages of using
monoclonal antibodies.
• Preparation of monoclonal antibodies.
• Applications of monoclonal antibodies.
MONOCLONAL IN E.COLI 2
4. WHAT ARE ANTIBODIES?
“ Every body has a specialized search and destroy army.
Antibodies are key players in that fight ”
Antibodies are protein made by plasma cells ( type of WBC’s) in
response to an antigen.
It is a Y shaped protein .
Every body naturally produces antibodies.
Are elements of the immune system produced by B lymphocytes.
Bind to foreign proteins in the body known as antigens, with the
aim of eliminating them.
Naturally circulate in the body searching for foreign bodies
(antigens).
MONOCLONAL IN E.COLI 4
5. MONOCLONAL IN E.COLI 5
Each tip of the “Y” of an antibody
contains a paratope (analogous to a
lock) that is specific for one particular
epitope (analogous to a key) on an
antigen, allowing these two structures
to bind together with precision.
They attach to the antigen and destroy
it by using various immune
mechanisms.
Classes- IgA, IgD, IgE, IgG, or IgM.
7. MONOCLONAL ANTIBODIES
Monoclonal: forming a clone which is derived asexually from a
single individual or cell.
Monoclonal antibodies are artificial antibodies that are produced
from a single clone of cells by fusing B lymphocytes to myeloma
cells.
The fusion of B-lymphocytes with myeloma cells by somatic cell
hybridization secretes desired antibody-producing elements which
are immortalized cell-lines known as a hybridoma.
These hybridomas produce homogenous monoclonal antibodies.
MONOCLONAL IN E.COLI 7
8. MONOCLONAL IN E.COLI 8
Monoclonal antibodies (mAbs) have the ability to recognize unique
binding sites (epitopes) found on the specific antigens.
This differentiates monoclonal antibodies from polyclonal
antibodies i.e. monoclonal antibodies are derived from a single B-
cell clone to target single epitopes, unlike polyclonal antibodies
that target multiple epitopes.
Monoclonal antibodies (mAbs) have been produced to target
receptors or other foreign proteins that are present on the surface
of normal cells and cancer cells.
10. TYPES OF MONOCLONAL ANTIBODIES
Accordingly, mAbs are of four broad types.
Murine: Made from mouse proteins, names of drugs based on this end in –
omab.
Chimeric: A combination of mouse and human proteins, names of drugs
based on this end in –ximab.
Humanized: Here small doses of mouse proteins are attached to human
proteins, names of drugs based on this end in –zumab.
Human: These are fully human proteins, names of drugs based on this end in
–umab.
MONOCLONAL IN E.COLI 10
12. ADVANTAGES OF USING MONOCLONAL ANTIBODIES
Monoclonal antibodies are one of the most successful
biotherapeutic drugs used in the treatment of many types of
cancer and autoimmune conditions.
They are also proven to reduce side-effects and improve patient
survival and well-being.
As side effects can be treated and reduced by using mice-human
hybrid cells or by using fractions of antibodies.
It is highly scalable, unlimited production source
It can produce antibodies when needed.
Antigen or immunogen need not be pure.
MONOCLONAL IN E.COLI 12
13. DISADVANTAGES OF USING MONOCLONAL ANTIBODIES
It is a time consuming project, it may take time between 6 months
to 9 months.
It is very expensive.
It needs considerable effort to produce them.
System is only well developed for mouse and rat and not for other
animals.
More than 99% of the cells do not survive during the fusion
process that reduces the range of useful antibodies that can be
produced against an antigen.
There is also a possibility of generating immunogenicity (the ability
of a molecule or substance to provoke an immune response).
MONOCLONAL IN E.COLI 13
15. STEPS IN PRODUCTION OF MONOCLONAL ANTIBODIES
MONOCLONAL IN E.COLI 15
Step 1-Immunization of rabbit or rat and extraction of B-
lymphocytes
In order to isolate B-lymphocyte producing certain antibodies,
rabbit or lab rat is immunized through repeated injection of
specific antigen (sheep RBCs)
A sample of B-cells is extracted from spleen of rabbit or rat.
16. MONOCLONAL IN E.COLI 16
Step 2- Fusion of myeloma cell with B- Lymphocytes
• The extracted B-lymphocytes is added to a culture of myeloma cell from bone
marrow.
Multiple myeloma cells are abnormal plasma cells (a type of white blood cell)
that build up in the bone marrow and form tumors in many bones of the body.
• Hybridoma cells formed by fusion of B-cell and myeloma cell.
• The fusion is done by using Polyethylene glycol (PEG) or by electroporation or
by using phages.
17. MONOCLONAL IN E.COLI 17
Step 3- Selection of hybridoma cell
The B-lymphocytes contains HPRT1 gene which codes for enzyme
Hypoxanthine-guanine phosphoribosyltransferase (HGPRT).
(HGPRT) is one of the central enzymes that recycle the building blocks of RNA and DNA
B- cells can grow in medium containing Hypoxanthine amonopterin thymine
(HAT media).
HAT Medium is a selection medium for mammalian cell culture.
Myeloma cell lack HPRT1 gene so, it does not produce HGPTR enzyme and it
does not grow in HAT medium.
The myeloma cell do not utilize Hypoxanthine.
Only hybridoma cell i.e.. fused cell between myeloma and B-cell can
survive and divide in HAT medium.
Screening is done to select hybridoma cells which are the desired cell for
monoclonal antibodies production
18. MONOCLONAL IN E.COLI 18
Step 4- Culture of Hybridoma cell.
The selected hybridoma cells are cultured in suitable medium like insulin,
transferon, ethanol, amine and other additional hormones.
Some commonly used culture media for hybridoma cell for production of
monoclonal antibodies are:
• DMEM (Dulbecco’s modified eagle medium)
• IMDM (Iscove’s Modified Dulbecco’s Medium)
• Ham’s F12
• RPMI 1640 medium (Roswell Park Memorial Institute
• 1640 medium)
19. MONOCLONAL IN E.COLI 19
Step 5-Inoculation of hybridoma cell into suitable host.
These hybridoma cells are then injected into lab animal so that they starts
to produce monoclonal antibodies.
These hybridoma cells may be frozen and store for future use.
20. MONOCLONAL IN E.COLI 20
Step 6-Extraction and purification of Monoclonal antibodies.
Monoclonal antibodies from host animal can be extracted and purified by the
following methods:
Ion exchange chromatography
Antigen affinity chromatography
Radial immunoassay
Immune precipitation
23. MONOCLONAL IN E.COLI 23
Escherichia coli
Escherichia coli, also known as E. coli, is a Gram-negative,
facultative anaerobic, rod-shaped bacteria.
It is commonly found in the animal feces, lower intestines
of mammals, and even on the edge of hot springs.
The simplicity and ease of fermentation has made E. coli
an ideal host for antibody fragment production.
E. coli advantages include:
• Well characterized genetics
• Short process development timeline
• Simple fermentation
• Scalability
• Less safety issues from viral contaminants
24. MONOCLONAL ANTIBODIES IN E.COLI
MONOCLONAL IN E.COLI 24
There are situations where E. coli may become the preferred production host
over the presently used mammalian cells.
A monoclonal antibody (mAb) was obtained from a mouse immunized with
solubilized outer membrane proteins extracted from a bovine
enterohemorrhagic strain of Escherichia coli (EHEC), O26.
EHEC is a pathogenic group of strains.
25. MONOCLONAL IN E.COLI 25
The mAb produced a strong immunoblot reaction
for E.coli 026.
This mAb was used in a sandwich enzyme-linked
immunosorbent assay (ELISA) format to screen
strains from animal and human sources, and all
reactive strains.
The antigen was detected in a group of strains
containing a high proportion of O26.
The association of the antigen detected by the MAb
with significant enteropathogenic E. coli and EHEC
virulence factors in isolates from both animal and
human enteric infections indicates a diagnostic
potential for the assay developed.
26. MONOCLONAL IN E.COLI 26
OBJECTIVE
The object was to produce monoclonal antibodies (mAbs) to EHEC surface
adhesion antigens, and to investigate their diagnostic application for the
detection of EHEC in animal and human enteric infections.
28. Disease diagnosis
• ELISA to test HIV, hepatitis, Herpes etc.
• RIA- to test viral infection.
• MAbs to Human chorionic gonadotropin.
Passive immunization or disease prevention
• Monoclonal antibodies based drugs can be used
to treat septic shock
• Used as vaccine
Detection and purification of biomolecules
• MAbs are very useful in determining the presence
and absence of specific proteins through western
blotting technique.
• Besides that, it can be used to classify strains of a
single pathogen. E.g. Neisseria gonorrhea can be
typed using Monoclonal antibodies.
MONOCLONAL IN E.COLI 28
29. SIDE-EFFECTS OF MONOCLONAL ANTIBODIES
MONOCLONAL IN E.COLI 29
Several monoclonal antibodies, such as bevacizumab and
cetuximab, can cause different kinds of side effects.
These side effects can be categorized into common and serious
side effects.
Common side effects include:
• Dizziness
• Headaches
• Allergies
• Fever
• Itching
• Insomnia
• Constipation etc.
30. 30
Serious side effects are
• Anaphylaxis
• Bleeding
• Arterial and venous blood clots
• Autoimmune thyroiditis
• Hypothyroidism
• Hepatitis
• Heart failure
• Cancer
31. PRESENTED BY
Thank You
Kashish Imran
-Basic concept and
introduction
Sadqua Urooj
-Advantages and
disadvantages of using
monoclonal antibodies.
Khadeeja Yasmeen
-Preparation of monoclonal
antibodies
Draksha
-Applications of monoclonal
antibodies.
MONOCLONAL IN E.COLI 31
-BSc Biotechnology, Sem II
PRESENTED TO
Dr. Saima Wajid|Professor
(Assistant)|Jamia Hamdard