Monoclonal antibodies (MAbs) are antibodies that are directed against a single antigen. They can be produced through hybridoma technology which involves fusing antibody-producing B cells with myeloma cells to form a hybrid cell line. This document outlines the process for producing MAbs including immunizing an animal, fusing B cells with myeloma cells, selecting antibody-producing hybridomas, cloning and mass producing the antibodies. MAbs have various diagnostic and therapeutic applications for diseases.
Monoclonal antibody
# Definition
# A general representation of the method used to produce monoclonal antibodies
#Applications
# Monoclonal antibodies for cancer.
#MAbs approved by the FDA for cancer include:
# Factor affecting monoclonal antibody immunogenicity
#Monoclonal Antibodies : 4 Types
# Monoclonal Antibodies : 4 Types images
Students of medical and allied subjects must be exposed to the concept of monoclonal antibodies for the efficient practice of clinical and laboratory medicine.
here is a powerpoint presentation on monoclonal antibodies fro students and researchers. if you are a student and looking for a presentation on the topic to present in class. this one is for you my friend.
An antibody (Ab), also known as an immunoglobulin (Ig), is a large, Y-shaped protein produced mainly by plasma cells that is used by the immune system to neutralize pathogens such as pathogenic bacteria and viruses.
Monoclonal antibodies are important reagents used in biomedical research, in diagnosis of diseases, and in treatment of such diseases as infections and cancer.
These antibodies are produced by cell lines or clones obtained from animals that have been immunized with the substance that is the subject of study.
Hybridoma Technology ( Production , Purification , and Application ) Sakshi Ghasle
Hybridoma technology revolutionized the field of immunology by enabling the production of monoclonal antibodies with high specificity and affinity. This presentation delves into the principles of DNA hybridoma technology, highlighting its significance in antibody production, therapeutic applications, and biomedical research. Learn about the key steps involved in generating hybridomas, from immunization to antibody screening, and discover the potential of recombinant DNA techniques in enhancing antibody engineering. Whether you're a student, researcher, or industry professional, this overview will provide valuable insights into the innovative world of hybridoma technology."
Uncover the wide-ranging applications of monoclonal antibodies in areas such as cancer therapy, autoimmune diseases, infectious diseases, and beyond. Learn about the latest advancements in antibody engineering and the development of novel therapeutic modalities, including bispecific antibodies, antibody-drug conjugates, and immune checkpoint inhibitors.
Whether you're a seasoned researcher or a newcomer to the field, this SlideShare presentation serves as a valuable resource for understanding the principles, techniques, and applications of hybridoma technology in modern biomedicine. Join a journey through the fascinating world of monoclonal antibodies and the groundbreaking science behind their creation.
Unlock the potential of hybridoma technology and propel your research to new heights. Dive into this SlideShare presentation now and explore the limitless possibilities of monoclonal antibody production with hybridoma technology.
Monoclonal antibody
# Definition
# A general representation of the method used to produce monoclonal antibodies
#Applications
# Monoclonal antibodies for cancer.
#MAbs approved by the FDA for cancer include:
# Factor affecting monoclonal antibody immunogenicity
#Monoclonal Antibodies : 4 Types
# Monoclonal Antibodies : 4 Types images
Students of medical and allied subjects must be exposed to the concept of monoclonal antibodies for the efficient practice of clinical and laboratory medicine.
here is a powerpoint presentation on monoclonal antibodies fro students and researchers. if you are a student and looking for a presentation on the topic to present in class. this one is for you my friend.
An antibody (Ab), also known as an immunoglobulin (Ig), is a large, Y-shaped protein produced mainly by plasma cells that is used by the immune system to neutralize pathogens such as pathogenic bacteria and viruses.
Monoclonal antibodies are important reagents used in biomedical research, in diagnosis of diseases, and in treatment of such diseases as infections and cancer.
These antibodies are produced by cell lines or clones obtained from animals that have been immunized with the substance that is the subject of study.
Hybridoma Technology ( Production , Purification , and Application ) Sakshi Ghasle
Hybridoma technology revolutionized the field of immunology by enabling the production of monoclonal antibodies with high specificity and affinity. This presentation delves into the principles of DNA hybridoma technology, highlighting its significance in antibody production, therapeutic applications, and biomedical research. Learn about the key steps involved in generating hybridomas, from immunization to antibody screening, and discover the potential of recombinant DNA techniques in enhancing antibody engineering. Whether you're a student, researcher, or industry professional, this overview will provide valuable insights into the innovative world of hybridoma technology."
Uncover the wide-ranging applications of monoclonal antibodies in areas such as cancer therapy, autoimmune diseases, infectious diseases, and beyond. Learn about the latest advancements in antibody engineering and the development of novel therapeutic modalities, including bispecific antibodies, antibody-drug conjugates, and immune checkpoint inhibitors.
Whether you're a seasoned researcher or a newcomer to the field, this SlideShare presentation serves as a valuable resource for understanding the principles, techniques, and applications of hybridoma technology in modern biomedicine. Join a journey through the fascinating world of monoclonal antibodies and the groundbreaking science behind their creation.
Unlock the potential of hybridoma technology and propel your research to new heights. Dive into this SlideShare presentation now and explore the limitless possibilities of monoclonal antibody production with hybridoma technology.
Hybridoma
Hybridomas are cells that have been engineered to produce a desired antibody in large amounts, to produce monoclonal antibodies.
Monoclonal antibodies can be produced in specialized cells through a technique now popularly known as hybridoma technology.
Hybridoma technology was discovered in 1975 by two scientists, G. Kohler and C. Milstein, were awarded Noble prize for physiology and medicine in 1984.
Production of Monoclonal Antibodies by Hybridoma Technology.pptxAnupkumar Sharma
The presentation includes the information about the production of monoclonal antibodies by hybridoma technology. The slides focus on the points like monoclonal and polyclonal antibodies, steps involved in hybridoma technology and its analytical, diagnostic, therapeutic and some miscellaneous applications. It also includes some marketed products of monoclonal antibodies.
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.
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Bio
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Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
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Slide 5: Plasmid Inheritance
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Slide 6: Mechanisms of Extrachromosomal Inheritance
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Slide 7: Examples of Extrachromosomal Inheritance
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Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
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Slide 9: Recent Research and Advances
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Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
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2. OVERVIEW :
INTRODUCTION
PRINCIPLE FOR
CREATION OF
HYBRIDOMA CELLS
STEPS INVOLVED IN
PRODUCTION OF
Mabs.
SECOND GENERATION
MAbs.
EXAMPLES
APPLICATIONS
DISADVANTAGES
ADVANTAGES
REFERENCES
3. INTRODUCTION :
• Monoclonal antibody is a single type of antibody that is directed against a specific
antigenic determinant(epitope).
• Natural source : Found in patients suffering from multiple myeloma.
• In 1975, George Kohler and Cesar Milstein achieved a large scale production of
MAbs. The pair won the 1984 Nobel Prize in Medicine and Physiology for this
discovery.
• The production of monoclonal antibodies by the hybrid cells is referred to as
Hybridoma Technology.
4. PRINCIPLE FOR CREATION OF HYBRIDOMA CELLS
• The mutated cells deficient in the enzyme HGPRT are grown in a medium
containing hypoxanthine, aminopterin and thymidine (HAT medium),
they cannot survive due to inhibition of de-novo synthesis of purine
nucleotides. Thus cells lacking the HGPRT enzyme, grown in HAT
medium die.
• The hybridoma cells possess the ability of myeloma cells to grow in vitro
with a functional HGPRT gene obtained from lymphocytes (with which
myeloma cells are fused). Thus, only the hybridoma cells can proliferate in
HAT medium and this procedure is used for selection.
5.
6. STEPS OF PRODUCTION OF
MONOCLONAL ANTIBODIES
1. Immunization
2. Cell fusion
3. Selection of hybridomas
4. Screening of products
5. Cloning and production
6. Characterization and storage.
7. IMMUNIZATION
• The animal is immunized using an appropriate antigen. The antigen along with an
adjuvant (mainly Freund’s complete) is injected subcutaneously.
• The injections are administered at multiple sites to increase the production of B-
lymphocytes, which are responding to the antigen.
• Three days prior to the sacrifice of the animal, a final dose of antigen is given
intravenously. This allows the synthesis of antibodies to it’s maximum level.
• The concentration of the desired antibodies is assayed in the serum of the animal at
frequent intervals during the course of immunization.
• When the serum concentration of the antibodies is optimal, the animal is sacrificed.
The spleen is aseptically removed and disrupted by mechanical or enzymatic
methods to release the cells.
• The lymphocytes of the spleen are separated from the rest of the cells by density
gradient centrifugation.
8. CELL FUSION
• The thoroughly washed lymphocytes are mixed with HGPRT defective
myeloma cells.
• The mixture of cells is exposed to polyethylene glycol (PEG) for a short
period of time.
• PEG is removed by washing and the cells are kept in a fresh medium.
• These cells are composed of a mixture of hybridomas (fused cells), free
myeloma cells and free lymphocytes.
9. SELECTION OF HYBRIDOMAS
• Within the 7-10 days of culture, only the hybridoma cells are able to grow
whereas the rest starts disappearing.
• Selection of a single antibody producing hybrid cells is very important. This
is possible if the hybridomas are isolated and grown individually.
• The suspension of hybridoma cells is so diluted that the individual aliquots
contain on an average one cell each. These cells, when grown in a regular
culture medium, produce the desired antibody.
10. SCREENING OF PRODUCTS
• The hybridomas must be screened for the secretion of the antibody of the
desired specificity.
• The two techniques namely ELISA and RIA are commonly used for this
purpose. In both the assays, the antibody binds to the specific antigen and
the unbound antibody and other components can be washed off.
• Thus, the hybridoma cells producing the desired antibody can be identified
by screening.
11. CLONING AND PROPAGATION
• The single hybrid cells producing the desired antibody are isolated and cloned.
• Two techniques are commonly employed for cloning hybrid cells-
i. Limiting dilution method
ii. Soft agar method
LIMITING DILUTION METHOD
• The suspension of hybridoma cells are serially diluted and the aliquots of each dilution are
put into microculture wells.
• The dilutions are so made that each aliquot in a well contains only a single hybrid cell.
• This ensures that the antibody produced is monoclonal.
SOFT AGAR METHOD
• The hybridoma cells are cultured in soft agar. It is possible to simultaneously grow many cells
in semisolid medium to form colonies. These colonies will be monoclonal in nature.
12. CHARACTERIZATION AND STORAGE
• The monoclonal antibody has to be subjected to biochemical and biophysical
characterization for the desired specificity.
• It is also important to elucidate the Mab for the immunoglobulin class or sub-
class, the epitope for which it is specific and the number of binding sites it
possesses.
13.
14.
15. SECOND GENERATION MONOCLONALANTIBODIES
• By employing site-directed mutagenesis, cysteine residues are introduced at
the predetermined positions on the Mab.
• These cysteine residues which facilitate the isotope labelling. This would be
more useful in diagnostic imaging and radioimmunotherapy.
16. ADVANTAGES
• Monoclonal antibodies truly represent a homogenous state of a single
molecular species.
• Each Mab is specific to a given antigenic determinant.
DISADVANTAGES
• Laborious and time-consuming.
• Since they are produced against a single antigenic determinant, therefore
they cannot differentiate the molecules well.
• Mab for human use should be totally free from all pathogenic organisms,
including viruses, but since there is no guarantee it poses a great danger.
17. APPLICATIONS
DIAGNOSTIC APPLICATIONS
Biochemical analysis for the diagnosis of pregnancy, cancers, hormonal disorders, infectious
diseases.
Diagnostic imaging for the detection of myocardial infarction, deep vein thrombosis, cancers,
atherosclerosis, bacterial infections.
THERAPEUTIC APPLICATIONS
Direct use as therapeutic agents to destroy disease causing organisms in the treatment of
cancers, AIDS, autoimmune diseases and organ transplantation.
As targeting agents in therapy as immunotoxins (for treatment of cancers) in drug delivery, for
dissolving blood clots in radioimmunotherapy.
PROTEIN PURIFICATION BY IMMUNOAFFINITY TECHNIQUES
MISCELLANEOUS APPLICATIONS INCLUDES CATALYTIC AGENTS LIKE ABZYMES,
USED IN AUTOANTIBODY FINGERPRINTING.
18.
19. REFERENCES :
• Textbook of Biotechnology by U. Satyanarayana. Edition-2013 ; Publisher-Books &
Allied Ltd.
• Textbook of Immunology (Kuby) by Judith.A. Owen, Jenni Punt, Sharon.A. Stanford
and Patricia.P.Jones. Edition-7th ; Publisher – W.H. Freeman and Company.
• www.Wikipedia.com
• www.genescript.com