Production and applications of monoclonal antibodiesKaayathri Devi
production and applications of monoclonal antibodies, monoclonal antibodies ,applications of monoclonal antibodies, production of monoclonal antibodies,
Production and applications of monoclonal antibodiesKaayathri Devi
production and applications of monoclonal antibodies, monoclonal antibodies ,applications of monoclonal antibodies, production of monoclonal antibodies,
Students of medical and allied subjects must be exposed to the concept of monoclonal antibodies for the efficient practice of clinical and laboratory medicine.
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 is a method for producing large number of identical antibodies called monoclonal antibodies.
It was discovered by G.kohler and C.milstein in 1975. they were awarded nobel prize for physiology and medicine in 1975.
The hybrid cells are produced by fusing B- lumphocyte with myeloma cells or tumour cells.
The B-lymphocyte have the ability to produce large number of antibodies and tumour cells have indefinite growth.
This is why two cells are used for the production of hybrid cell
Hybridoma technology is a method for producing large numbers of identical antibodies (also called monoclonal antibodies). This process starts by injecting a mouse (or other mammals) with an antigen that provokes an immune response.
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.
INTRODUCTION: Monoclonal antibodies can be produced through a technique known as hybridoma technology.
HISTORY: The production of monoclonal antibodies was invented by Niels K.J. Georges, J.F. Kohler and Cesar Milstein in 1975.
PRINCIPLE FOR CREATION OF HYBRIDOMA CELLS: HAT (hypoxanthine aminopterin and thymidine) medium – Only hybridoma cells can proliferate in HAT medium.
PRODUCTION OF MONOCLONAL ANTIBODIES (HYBRIDOMA TECHNOLOGY): The establishment of hybridomas and production of monoclonal antibodies involves the following steps-
Immunization (ii) Cell fusion (iii) Selection of hybridomas (iv) Screening the products (v) Cloning and propagation (vi) Characterization and storage.
ADVANTAGES AND DISADVANTAGES OF MONOCLONAL ANTIBODIES:
Advantages- Monoclonal antibodies is specific to a given antigenic determinant.
Disadvantages- There is no guarantee that monoclonal antibodies produced is totally virus-free, despite the purification.
APPLICATIONS OF MONOCLONAL ANTIBODIES: Diagnostic applications, therapeutic applications, protein purification and miscellaneous applications.
REFERENCES:
• Satyanarayana, U. 2016. Biotechnology. Books and Allied (P) Ltd, Kolkata. pp. 213-226.
• Gupta, P.K. 2016. Biotechnology and Genomics. Rastogi Publications, Meerut. pp. 299-311.
• Owen, J.A., Punt J., Stranford, S.A. and Patricia, P.J. 2013. Kuby Immunology. 7th Ed. W.H. Freeman and Company, New York. pp.645-655.
• Singh, B.D. 2017. Biotechnology Expanding Horizons. Kalyani Publishers, New Delhi. pp. 172-174.
• Dubey, R.C. and Maheshwari, D.K. 2018. A Textbook of Microbiology. S Chand and Company Limited, New Delhi. pp. 662-663.
Monoclonal antibodies drug targeting particuler carrier systemRoshan Lal Singh
monoclonal antibodies drug targeting particulate carrier system
presented by : - Roshan Lal Singh student of M.pharma 2nd semester University institute of pharmacy Pt. R. S. U. Raipur (C.G.).
Students of medical and allied subjects must be exposed to the concept of monoclonal antibodies for the efficient practice of clinical and laboratory medicine.
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 is a method for producing large number of identical antibodies called monoclonal antibodies.
It was discovered by G.kohler and C.milstein in 1975. they were awarded nobel prize for physiology and medicine in 1975.
The hybrid cells are produced by fusing B- lumphocyte with myeloma cells or tumour cells.
The B-lymphocyte have the ability to produce large number of antibodies and tumour cells have indefinite growth.
This is why two cells are used for the production of hybrid cell
Hybridoma technology is a method for producing large numbers of identical antibodies (also called monoclonal antibodies). This process starts by injecting a mouse (or other mammals) with an antigen that provokes an immune response.
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.
INTRODUCTION: Monoclonal antibodies can be produced through a technique known as hybridoma technology.
HISTORY: The production of monoclonal antibodies was invented by Niels K.J. Georges, J.F. Kohler and Cesar Milstein in 1975.
PRINCIPLE FOR CREATION OF HYBRIDOMA CELLS: HAT (hypoxanthine aminopterin and thymidine) medium – Only hybridoma cells can proliferate in HAT medium.
PRODUCTION OF MONOCLONAL ANTIBODIES (HYBRIDOMA TECHNOLOGY): The establishment of hybridomas and production of monoclonal antibodies involves the following steps-
Immunization (ii) Cell fusion (iii) Selection of hybridomas (iv) Screening the products (v) Cloning and propagation (vi) Characterization and storage.
ADVANTAGES AND DISADVANTAGES OF MONOCLONAL ANTIBODIES:
Advantages- Monoclonal antibodies is specific to a given antigenic determinant.
Disadvantages- There is no guarantee that monoclonal antibodies produced is totally virus-free, despite the purification.
APPLICATIONS OF MONOCLONAL ANTIBODIES: Diagnostic applications, therapeutic applications, protein purification and miscellaneous applications.
REFERENCES:
• Satyanarayana, U. 2016. Biotechnology. Books and Allied (P) Ltd, Kolkata. pp. 213-226.
• Gupta, P.K. 2016. Biotechnology and Genomics. Rastogi Publications, Meerut. pp. 299-311.
• Owen, J.A., Punt J., Stranford, S.A. and Patricia, P.J. 2013. Kuby Immunology. 7th Ed. W.H. Freeman and Company, New York. pp.645-655.
• Singh, B.D. 2017. Biotechnology Expanding Horizons. Kalyani Publishers, New Delhi. pp. 172-174.
• Dubey, R.C. and Maheshwari, D.K. 2018. A Textbook of Microbiology. S Chand and Company Limited, New Delhi. pp. 662-663.
Monoclonal antibodies drug targeting particuler carrier systemRoshan Lal Singh
monoclonal antibodies drug targeting particulate carrier system
presented by : - Roshan Lal Singh student of M.pharma 2nd semester University institute of pharmacy Pt. R. S. U. Raipur (C.G.).
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.
Hybridoma technology is a method for producing large numbers of identical antibodies (also called monoclonal antibodies). This process starts by injecting a mouse (or other mammal) with an antigen that provokes an immune response.
Hybridoma technology is a method for producing large numbers of identical antibodies (also called monoclonal antibodies). This process starts by injecting a mouse (or other mammal) with an antigen that provokes an immune response.
These lecture slides, by Dr Sidra Arshad, offer a quick overview of physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Ve...kevinkariuki227
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Anti ulcer drugs and their Advance pharmacology ||
Anti-ulcer drugs are medications used to prevent and treat ulcers in the stomach and upper part of the small intestine (duodenal ulcers). These ulcers are often caused by an imbalance between stomach acid and the mucosal lining, which protects the stomach lining.
||Scope: Overview of various classes of anti-ulcer drugs, their mechanisms of action, indications, side effects, and clinical considerations.
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
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These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
MANAGEMENT OF ATRIOVENTRICULAR CONDUCTION BLOCK.pdfJim Jacob Roy
Cardiac conduction defects can occur due to various causes.
Atrioventricular conduction blocks ( AV blocks ) are classified into 3 types.
This document describes the acute management of AV block.
2. Monoclonal antibodies
An antibody is a protein used by the immune system to identify and
neutralize foreign objects like bacteria and viruses. Each antibody
recognizes a specific antigen unique to its target.
Monoclonal antibodies (mAb) are antibodies that are identical because
they were produced by one type of immune cell, all clones of a single parent
cell
Characters of Monoclonal Antibodies
Monoclonal antibodies (mAb) are a single type of antibody that are
identical and are directed against a specific epitope (antigen, antigenic
determinant) and are produced by B-cell clones of a single parent or a
single hybridoma cell line.
A hybridoma cell line is formed by the fusion of a one B-cell lymphocyte
with a myeloma cell.
3. Principle of Hybridoma technology for mAB production
inject the protein into a
mouse.
remove the spleen.
identify cells producing
antibodies.
separate these cells and grow
in tissue culture tubes.
screen each Ab for cross
reactivity.
select the Ab which doesn't
cross react with any other
protein.
Fusion agent:
Polyethylene glycol
Medium:
HAT Medium {Hypoxathine-
Aminopterin-Thymidine}
4. Production of mAB by Hybridoma technology
Four basic steps are involved:
1)Immunization
2)Generation of B cells hybridomas by fusing prime b cells and
myeloma cells
3)Selection and screening of resulting clones
4)Cloning by propagating the desired hybridoma
5.
6. Steps: Immunization
• Immunization of animal with immunogen (mixed with an
adjuvant like Al salts, Freund’s complete or incomplete
adjuvant
• The serum of animals is assayed for the relative
concentration, if found to be normal, animal is sacrificed
and spleen which contains the large number of plasma
cells is dissociated into single splenocytes by mechanical
or enzymatic method
7. Steps: Cell fusion
• Splenocytes are mixed with myeloma cells (HGPRT) in an
appropriate medium and is exposed to high concentration of PEG for
short period and fusion is allowed to proceed
• The use of HGPRT cells (that cannot grow in HAT medium) assured
that only hybridomas (hybrid myeloma spleen cells) are selected
• HGPRT, Ab-
myeloma cell was chosen as ideal fusion partner. This
fusion partner has the immortal properties of a cancer cell but does
not secrete its antibody or gene product. Hybridomas generated with
this fusion partner thus secrete only the antibody from the B cell
partner.
• These hybridomas can be propagated in tissue culture to give rise to
large clones secreting homogenous monoclonal antibodies
8. Steps: Selection and screening of
the clones for the monoclonal
antibody specificity
• HAT medium allow only hybridomas cells to grow
• They are removed from the tissue culture flasks, and transferred to
regular culture medium and aliquots are distributed among the well of
96-well plastic culture plates.
• The supernatant of each hybridoma culture can be assayed for a
particular antigen specificity by two methods, i.e., ELISA or RIA
9. Steps: Cloning of hybridomas
secreting specific monoclonal
antibody
• Single cells secreting the desired antibody are isolated from positive
cultures and propagated into cell lines. Two cloning techniques are most
widely employed:
• Limiting dilution (Cells in a culture are enumerated, diluted and aliquoted
into new wells so that only one cell found in any well. Cells are allowed to
regrow and the procedure is repeated several times to increase the
probability that all cells in a given culture are monoclonal)
• Soft agar (many malignant cells may grow, forming spherical colonies, in a
semisolid medium containing low amounts of agar. If the culture can be
reliably dispersed into single cells and the cell concentration is such that
the colonies will be well spaced, then visible colonies picked out of the
agar are likely to be monoclonal
10. Applications of mAB: Diagnostic
1) MAbs in pregnancy testing
Beta HCG ABs
Immunoprecipitation (Micral test): It is
based on the colour shift of monoclonal
antibody to human albumin labelled with
gold
11. 4) mABs IN IMMUNODIAGNOSTIC TESTS
Monoclonal antibodies can also be used
to purify a substance with techniques
called immuno-precipitation and
affinity chromatography.
12. Application of mABs: Therapeutic
1)General
Radioisotope immunoconjugates
mABs conjugates deliver cytotoxic doses of radioactivity to target cells.
Choice of isotopes is critical. Some currently evaluated isotopes are
.Most studies are used β emitters. mABs carrying these emitters are
able to kill nearby Stauder’s cells which cause damage to nearby non-
target cells.
Toxin drug conjugate
mABs are being evaluated for the delivery of potent toxin and drugs to
target cells while reducing toxicity to non- target tissues. Methods
involved is an exclusion of intrinsic infectivity and incorporation of
selectivity for presentation to target cells
2) Transplantation
Organ
Bone marrow
Acute rejection of organ transplants by inhibiting Interleukin-2 on
activated T cells
13. 3) Infectious disease
Micro-organism
mABs against micro-organism can be readily made. mABs can be
used for identification, isolation, detection, structural investigation and
genetic variation and classification of micro-organism.
Parasites
Most important among this class is the malarial parasite. vaccination
against this widespread disease is difficult due to complex life cycle of
these organism. mABs when available will help in treatment,
interception and inactivation pd the parasite life cycle
viruses Bacteria
Influenza Streptococcus A
Rabies Streptococcus B
Hepatitis B Neisseria gonorrhoeae
Herpes Simplex Pseudomonas aeruginosa
Epstein- Barr M.tuberculosis
14. 4) Cardiovascular disease
A murine Mab, 7E3, specific for glycoprotein II b/Iia fibrinogen receptor of
platelets has been evaluated by Centocor
5) Autoimmune disease
mAB directed against B and T lymphocytes are currently being evaluated for
autoimmune diseases.
6) Cancer
1)Radioimmunotherapy (RIT):
Involves the use of radioactively conjugated murine antibodies against
cellular antigens to limit radiation exposure.
Murine antibodies were especially chosen, as their high immunogenicity
promotes rapid clearance from the body.
Ex:Tositumomab in non-Hodgkins lymphoma
2)Antibody-directed enzyme prodrug therapy (ADEPT)
It involves the application of cancer associated monoclonal antibodies which
are linked to a drug-activating enzyme.
Subsequent systemic administration of a non-toxic agent results in its
conversion to a toxic drug, and resulting in a cytotoxic effect which can be
targeted at malignant cells
15. 3)Immunoliposomes: are antibody-conjugated liposomes.
Liposomes - carry drugs or therapeutic nucleotides - conjugated with
monoclonal antibodies - directed against malignant cells.
Tissue-specific gene delivery using immuno-liposomes has also been
achieved in brain, and breast cancer tissue
16. 1) Lymphocyte phenotyping
mABs specific for membrane proteins can be applied to identify various
stages of lymphocyte differentiation. Eg. T helper cell express CD8
membrane protein in both human and mice.
Purification of protein
mABs can be made to any minor protein (P) in a complex mixture since
any Hybridoma clone that secretes antibody to proteins other than P are
eliminated during the screening phase of monoclonal production. This
mABs to a particular protein can be used for its purification
Radioimmunoassay
mABs have the potential to replace or supplement conventional antibodies
in a series of assay. mABs reacts with different sites of single molecules
allows to produce highly specific radio-assays in which one of mABs is
Application of mABs: Analytical