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.
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.
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.
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
What are antibodies?
An antibody is a protein used by immune system to identify and neutralize foreign agents like bacteria and viruses.
Each antibody recognizes a specific antigen unique to its target.
BIOTECHNOLOGY IS
CHALLENGING SUBJECT TO TEACH AND UNDERSTAND ......
ITS A VERY INTERESTING TO LEARN ABOUT HYBRIDOMA TECHNOLOGY .. THEIR PRODUCTION AND
APPLICATION ALSO ....
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
What are antibodies?
An antibody is a protein used by immune system to identify and neutralize foreign agents like bacteria and viruses.
Each antibody recognizes a specific antigen unique to its target.
BIOTECHNOLOGY IS
CHALLENGING SUBJECT TO TEACH AND UNDERSTAND ......
ITS A VERY INTERESTING TO LEARN ABOUT HYBRIDOMA TECHNOLOGY .. THEIR PRODUCTION AND
APPLICATION ALSO ....
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Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
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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
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Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
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Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
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MCAB
1.
2. Antibodies
An antibody (Ab), also known as an immunoglobulin
(Ig), is a large Y-shaped protein produced by B cells
that is used by the immune system to identify and
neutralize foreign objects such as bacteria and viruses.
“An antibody produced by a single clone of cells.
Monoclonal antibody is therefore a single pure
type of antibody”
Each monoclonal antibody recognizes a specific
antigen.
3.
4.
5. ANTIBODIES
POLYCLONAL MONOCLONAL
Derived from different B
Lymphocytes cell lines.
Batch to Batch variation
affecting Ab reactivity &
treatment.
NOT Powerful tools for
clinical diagnostic tests.
Derived from a single B cell
clone.
No Batch to Batch
variations. Effectiveness of
Ab is much more
predictable.
Enable the development of
secure immunoassay
systems.
6.
7. The types of mAb designed
Murine source mAbs: rodent mAbs with excellent affinities
and specificities generated using conventional hydrioma
technology. Clinical efficacy compromised by HAMA(human
anti murine antibody) response, which lead to allergic or
immune complex hypersensitivities.
Chimeric mAbs: chimers combine the human constant
regions with the intact rodent variable regions. Affinity and
specificity unchanged. Also cause human antichimeric
antibody response (30% murine resource)
Humanized mAbs: contained only the CDRs of the rodent
variable region grafted onto human variable region
framework
8. 1) Homogeneity:
Monoclonal antibody represents a single antibody molecule that
binds to antigens with the same affinity and promote the same
effectors functions.
2) Specificity:
The product of a single hybridoma reacts with the same epitope on
antigens.
3) Immunizing Antigen:
Need not be pure or characterized and is ultimately not needed to
produce large quantities of antibody.
4) Selection:
It is possible to select for specific epitope specificities and generate
antibodies against a wider range of antigenic determinants.
5) Antibody Production:
Unlimited quantities of a single well-defined monospecific reagent.
9. 1) Affinity:
Average affinity of monoclonal antibodies are generally
lower than polyclonal antibodies.
2) Effector Functions:
Because antibody is monoclonal, it may not produce the
desired biologic response.
3) Specificity:
Monoclonals against conformational epitopes on native
proteins may lose reactivity with antigens.
4) Cross reactions:
Antibodies sometimes display unexpected crossreactions
with unrelated antigens.
5) Time and effort commitment:VERY LARGE.
10.
11.
12. Process by which bulk quantities of targeted antibodies
against a specific antigen are produced.
Monoclonal antibodies are produced via multiple/identical
copies of a certain cell called a hybridoma.
To create Hybridoma cells the fusion of 2 cells are needed in
order to combine the characteristics of the 2 cells into 1 cell.
1 of the cells is a producing cell antibody which is a B-
Lymphocyte used from a laboratory mouse and the other is a
tumor cell named myeloma.
Tumor cells have the ability to grow indefinitely and at an
exceeding rate from normal cell growth.
Laboratory produced Hybridoma cells replicate much faster
than normal antibody producing cells, and the individual
hybridomas produce the specific antibodies for an indefinite
period of time.
13. Hybridoma cells manufacture the specific monoclonal
antibody that was originally produced by the B-
Lymphocyte cell.
The original B-Lymphocyte cell will produce the
Monoclonal antibody depending on the kind of antigen
that was injected into the mouse just prior to the
harvesting of the B-Lymphocyte cells.
Eg: If the mice were injected with a certain virus, the
mouse will have B-Lymphocytes that produce those
specific virual antibodies. Fusion with a tumor cell to
make the hybridoma, result in the production of
monoclonal antibodies against the specific virus.
14.
15. The hybridoma cells are placed into media that can help
them grow and produce the bulk quantities of monoclonal
antibodies.
There are 2 ways for growing monoclonal antibodies, 1 is
to grow them in laboratory flasks meaning In Vitro, and
the other is to grow them in the stomach lining of mice.
Injecting the hybridomas into the mice is the familiar
method of harvesting monoclonal antibodies. This
method is done by mixing spleen cells from the mouse
that has been immunized with the desired antigen with
myeloma cells. The myeloma cells need to have lost their
ability to synthesize HGPRT enzyme (hypoxanthine-
guanine-phosphoribosyltransferase) which enables the
cells to synthesize purines using an extracellular source of
hypoxanthine as a precursor.
16. Cells have another pathway that they synthesize
purines so lack of HGPRT is not a problem for the cell
but when cells are exposed to aminopterin they are
unable to use this other pathway and are fully
dependent on HGPRT for their survival. So to
summarize, unfused myeloma cells can’t grow since
they lack HGPRT and unfused normal spleen cells
can’t grow since they have a limited life-span.
Hybridoma cells are grow indefinitely since the spleen-
cell copartner supplies HGPRT and the myeloma
partner is immortal.
17. The first step is transferring of the cell fusion mixture to HAT culture
medium which contains hypoxanthine, aminopterin & pyrimidine
thymidine.
The 2nd step is testing the supernatants from each culture in order to
locate the producing the desired antibody.
One must isolate the single cells from each antibody-positive culture
and subculture them, this represents the clone which its antibodies
are monoclonal.
Every single cell culture secretes a specific kind of antibody that is
directed against a certain determinant/selected antigen.
The 3rd step is scaling up the size of the cultures of the successful
clones.
Hybridoma cultures can be grown indefinitely in vitro in culture
vessels which yield 15-65 µg/ml and in vivo using mouse, where the
antibody concentration in the serum 0.5-15 mg/ml.
In the past years, animal welfare activists in worldwide are trying to
limit the use of mice for the production of monoclonal antibodies.
When the monoclonal antibody is produced it can be used as a
probe to track down, bind to and purify the specific protein that
induced its formation.
18. 1) FILTRATION METHOD:
Cells, cell debris,
lipids, and clotted
material are first
removed, typically by
filtration with a 0.45
um filter.
19. a) Most of the charged impurities are usually
anions such as nucleic acids and endotoxins.
These are often separated by ion exchange
chromatography.
b) column chromatography can also use which is
much quicker method.
22. Monoclonal antibodies used for autoimmune
diseases include infliximab and adalimumab,
which are effective in rheumatoid
arthritis, Crohn's disease and ulcerative Colitis.
Basiliximab and daclizumab activated T cells and
thereby help preventing acute rejection of kidney
transplants.
Omalizumab inhibits human immunoglobulin E
(IgE) and is useful in moderate-to-severe
allergic asthma.
23. Antibodies are used in several diagnostic tests to
detect small amounts of drugs, toxins or
hormones.
e.g. monoclonal antibodies to human chorionic
gonadotropin (HCG) are used in pregnancy test
kits.
Another diagnostic uses of antibodies is the
diagnosis of AIDS by the ELISA test.
24.
25. Antibody Brand name Type Indication
Abciximab ReoPro chimeric Cardiovascular disease
Adalimumab Humira human Several auto-immune
disorders
Alemtuzumab Campath humanized Chronic lymphocytic
leukemia
Basiliximab Simulect chimeric Transplant rejection
Belimumab Benlysta human Systemic lupus
erythematosus
Bevacizumab Avastin humanized
Colorectal cancer, Age
related macular
degeneration
Brentuximab
Vedotin
Adcetris Chimeric Hodgkin lymphoma
26. Antibody Brand name Type Indication
Canakinumab Ilaris Human
Cetuximab Erbitux chimeric Colorectal cancer, Head
and neck cancer
Certolizumab
pegol Cimzia humanized Crohn's disease
Daclizumab Zenapax humanized Transplant rejection
Denosumab Prolia , Xgeva Human Postmenopausal
osteoporosis
Eculizumab Soliris humanized Paroxysmal nocturnal
hemoglobinuria
Efalizumab Raptiva humanized Psoriasis
27. Antibody Brand name Type Indication
Palivizumab Synagis humanized Respiratory Syncytial
Virus
Panitumumab Vectibix human Colorectal cancer
Ranibizumab Lucentis humanized Macular degeneration
Rituximab Rituxan,
Mabthera chimeric Non-Hodgkin
lymphoma
Tocilizumab Actemra Humanised Rheumatoid arthritis
Tositumomab Bexxar murine Non-Hodgkin
lymphoma
Trastuzumab Herceptin humanized Breast cancer