This document provides information about monoclonal antibodies including their production, types, and applications. It discusses how monoclonal antibodies are produced using the hybridoma technique which fuses antibody producing B cells with myeloma cells. This results in immortal cell lines that produce identical antibodies targeting a single epitope. The document contrasts monoclonal and polyclonal antibodies and describes the evolution of monoclonal antibodies from murine to humanized and human forms to reduce immunogenicity. It also covers monoclonal antibody nomenclature, pharmacokinetics, adverse effects, and therapeutic uses.
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.
Production and applications of monoclonal antibodiesKaayathri Devi
production and applications of monoclonal antibodies, monoclonal antibodies ,applications of monoclonal antibodies, production of monoclonal antibodies,
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.
Production and applications of monoclonal antibodiesKaayathri Devi
production and applications of monoclonal antibodies, monoclonal antibodies ,applications of monoclonal antibodies, production of monoclonal antibodies,
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 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
Introduction, causes and symptoms, Mechanism and treatment are been explained about this deadly disease SCID where production of T and B cells is affected.
It includes general introduction to antibodies; Monoclonal antibodies; comparison between Polyclonal & Monoclonal antibodies; Hybridoma Technology & Hyridoma Selection; advantages & disadvantages of mABs; Applications of mABs; Recombinant Monoclonal antibodies production through Antibody Engineering.
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 ....
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
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
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
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
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
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.
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
New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
STATEMENT OF NEED
Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
Target Audience
Hematology/oncology fellows, attending faculty, and other health care professionals involved in the treatment of patients with mantle cell lymphoma (MCL).
Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
2.) Evaluate emerging data on targeted therapeutic approaches for treatment-naive and relapsed/refractory MCL and their applicability to older adults
3.) Assess mechanisms of resistance to targeted therapies for MCL and their implications for treatment selection
3. Seminar layout
Antibody
Polyclonal Antibody
Monoclonal Antibody
Hybridoma technique
Production of Monoclonal Antibody
Evolution of Monoclonal Antibody
Nomenclature of Monoclonal Antibody
Types of Monoclonal Antibody
Pharmacokinetics & Adverse Effects
Therapeutic Potentials of Monoclonal Antibody
4. What are 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.
Polyclonal antibodies are antibodies that are
derived from different cell lines. They differ in
amino acid sequence.
Immunoglobulin (Ig) are structurally related
glycoproteins that function as antibodies
5. The Structure of an
Antibody
2 identical light chains (~220 amino
acids long)
Variable domain: VL
Constant domain: CL
2 identical heavy chains (~440 amino
acids long)
Variable domain: VH
3 Constant domains: CH1, CH2, CH3
Covalent, disulfide bonds between
cysteine residues
Flexible “hinge region”
6. Antibodies have two major functions:
• Recognize and bind antigen
• Induce immune responses after binding
The variable region mediates binding
• Affinity for a given antigen is
determined by the variable region
• The variable region confers absolute
specificity for an antigen
The constant region mediates immune
response after binding
• Different classes of constant regions
generate different isotypes
• Different isotypes of antibody have
differing properties
Antibody Function
Constant
region
Variable
region
7. Antibodies as Drugs
Antibodies are naturally occurring
Discovery of their innate properties hinted at
great therapeutic potential
High-specificity in binding
Already present in the body
Can activate and couple components of the immune
system
Modification to structure and refinement in
production methods have made antibodies a
viable modern drug
8. History
1975 :
Hybridoma Technology
George Kohler and Cesar
Milstein devised a method to
obtain large amounts of a mAb
1988 :
The Nobel Prize for Medicine
- In 1988, Greg Winter et al
pioneered the techniques to
humanize monoclonal
antibodies
9. History
1986 first monoclonal antibody reached
the market – Muromonab-CD3
2003 First fully human monoclonal
antibody – Adalimumab
10. Polyclonal antibodies are a mixture of antibodies with
different antigen binding sites that may bind to different
epitopes or antigens of the immunizing agent with
varying affinities.
Produced by immunizing an animal with the appropriate
antigen - wide array of B cells will be stimulated to
produce anti-protein antibodies.
Antibodies may be made to a number of different
epitopes of the protein.
Even antibodies that bind to the same epitope may have
different antigen-binding sites and bind the epitope with
different affinity.
Polyclonal Antibodies
11. The serum obtained from an immunized animal is
referred to as a polyclonal antiserum.
Contains antibody to different epitopes and different
antigens that were present in the immunizing inoculum
The immunized animal’s serum is collected.
Antibodies can then be purified from the serum.
Since one antigen induces the production of many
antibodies the result is a ‘polyclonal’ mixture of
antibodies.
ATG ( Anti thymocyte globulin)
ALG ( Anti lymphocyte globulin, Lymphocyte immune
globulin)
12. Monoclonal Antibodies
• A class of highly specific antibodies produced by the
clones of a single hybrid cell
• Produced by fusing a B cell secreting the desired
antibody with a tumour cell (myeloma cell) capable of
growing indefinitely
• Fused cell called hybridoma
• Monoclonal antibodies all have identical antigen-
binding sites
• bind to the same epitope with same affinity
• same antibody class (isotype)
13. Antibodies
Polyclonal Monoclonal
Produced by: Many B cell clones A single B cell
clone
Bind to: Multiple epitopes
of all antigens
used in the
immunization
A single epitope of
a single antigen
Antibody
class:
A mixture of
different Ab
classes (isotypes)
All of a single Ab
class
Ag-binding
sites:
different antigen-
binding sites
All Abs have the
same antigen
binding site
14. Antibodies
Polyclonal Monoclonal
Cost Less expensive More expensive
Yield Limited supply Infinite supply
Ease Easily, rapidly
produced
Time
consuming,
more technical
skill
Potential for
cross-
reactivity
High Low
15.
16.
17. PRODUCTION OF MONOCLONAL ANTIBODY
Step 1: - Immunization Of Mice & Selection Of Mouse
Donor For Generation Of Hybridoma cells
HYBRIDOMA TECHNOLOGY
ANTIGEN ( Intact cell/
Whole cell membrane/
micro-organisms ) +
ADJUVANT
(emulsification)
Ab titre reached in Serum
Spleen removed
(source of cells)
18. PRODUCTION OF MONOCLONAL ANTIBODY
Step 2: - Screening Of Mice For Antibody Production
HYBRIDOMA TECHNOLOGY
After several
weeks of
immunization
Serum Antibody Titre Determined
(Technique: - ELISA / Flow cytometery)
Titre too low
BOOST
(Pure antigen)
Titre High
BOOST
(Pure antigen)
2 weeks
19. PRODUCTION OF MONOCLONAL ANTIBODY
Step 3: - Preparation of Myeloma Cells
HYBRIDOMA TECHNOLOGY
Immortal Tumor Of Lymphocytes
+ 8 - Azaguanine
Myeloma Cells
High Viability & Rapid Growth
HGPRT-
Myeloma Cells
20. PRODUCTION OF MONOCLONAL ANTIBODY
Step 4: - Fusion of Myeloma Cells with Immune Spleen Cells
&
Selection of Hybridoma Cells
HYBRIDOMA TECHNOLOGY
FUSION
PEG
MYELOMA CELLSSPLEEN CELLS
HYBRIDOMA CELLS
ELISA PLATE
Feeder Cells
Growth Medium
HAT Medium
1. Plating of Cells in
HAT selective
Medium
2. Scanning of Viable
Hybridomas
21. PRODUCTION OF MONOCLONAL ANTIBODY
Step 4: - Cloning of Hybridoma Cell Lines by “ Limiting
Dilution” or Expansion
HYBRIDOMA TECHNOLOGY
A. Clone Each +ve Culture
B. Test Each Supernatant for Antibodies
C. Expand +ve Clones
Mouse
Ascites
Method
Tissue
Culture
Method
22. Myeloma cells have been genetically
engineered such that they can not use
hypoxanthine, aminopterin, and thymidine
(HAT medium) as a source for nucleic acid
biosynthesis and will die in culture (lack
HGPRT enzyme)
Spleen cells (B cells) have limited life span
Only B cells that have fused with the
engineered myeloma cells will survive in
culture when grown in HAT medium.
Hybridoma Selection The “HAT Trick”
23.
24. Conventional production of mAbs
The hybridoma technology:
spleen cells from immunized mice are fused with the murine myeloma cells.
The several process had been developed at large scale.
According to the different cell culture methods, it can calisifed into four fields
1. Robottle cell culture process.
2. Membrane binded cell culture process
3. Microcarrier cell culture process
4. Suspended cell culture process
25. Production in animals ( In-vivo )
Mouse ascites method
•Hybridoma cells injected in mouse
•Produce ascites
•Fluid contains high concentration of
Ab’s
•No further concentration required
•Purification required
•Easy and inexpensive
•Animal mortality
26. Ethical issues:
Freund’s complete adjuvant (FCA) (to enhance the immune
response): painful lesions at the injection site.
Pristane as a "priming" agent - granulomatous reactions
Respiratory distress: due to ascites.
Shock - rapid fluid loss
FCA and pristane should not be used where it is possible to
use no adjuvant or less irritant adjuvants.
FCA should not be used more than once in individual mice.
The volume of FCA and pristane used should not exceed
0.1ml and 0.2ml respectively.
Individual mice should not be inoculated with adjuvant more
than 3 times.
A priming agent should not be used in individual mice more
than once.
Ascites fluid should only be harvested once at the time of
euthanasia.
27. Production in cell culture ( In-vitro)
Batch tissue culture method:
• Grow hybridoma cells in batches
• Purify Mabs from the culture media
• Fetal bovine serum commonly used
• low concentration
• denaturation during concentration
Semi permeable membrane based system :
• A barrier – hollow fibre or a membrane
• Larger compartment containing culture media
• Smaller chamber to isolate cells and Mabs
• High concentrations
• Method of choice for large scale production
28. Origin
First generation
Murine, rabbit or rat proteins purified after
immunisation with antigen
Abs to these proteins (Ag) generated in
patients: human antimurine antibody (HAMA)
Block effectiveness of therapy
Adverse events serum sickness or anaphylaxis
29. Origin
Second generation
DNA technology or genetic engineering used
to construct hybrids composed of human Abs
regions with murine
Chimeric Abs
Humanized
Human
30. EVOLUTION OF MONOCLONAL ANTIBODY
1. TRANSGENIC
2. LIBRARIES
a.BACTERIOPHAGE
b. mRNA
c. Cell Surface
Ist
generation
mab
2nd generation mab
Daclizumab
33. Murine
Derived from mice
Patients treated with murine mAbs develop a
human antimouse antibody (HAMA) response
Rapid clearance of the mAb
Poor tumour penetration
Hypersensitivity reactions
90Y-ibritumomab
131I -Tositumomab
34. Chimeric Abs
Antigen binding parts (variable region) of mouse
with effector parts (constant region) of human
Infliximab
Abciximab
Rituximab
35. Humanized
Human Ab with complimentary determining region
(CDR) or hypervariable region from non human
source
Daclizumab
Trastuzumab
36. Human Abs
Recombinant DNA technology:
Genes for variable Fab portion of human Abs
is inserted in genome of bacteriophages &
replicated
Mixed with Ag & complementary Ab
producing phages selected
e.g. Adalimumab
38. Human Abs
Even human Ab can provoke anti-idiotype
responses
HACA (human antichimera antibodies)
HAHA (human antihuman antibodies)
Plant Derived Mabs
- Plant Genetic Engineering
- Transgenic Plants
- Transgenic tobacco, soyabeen, alfalfa etc.
Lack of animal pathogenic contaminants
Low production cost
Ease of agricultural scale up
41. Examples
ab- + -ci- + -xi- + -mab: chimeric
monoclonal antibody used on the
cardiovascular system.
tras- + -tu- + -zu- + -mab:
humanized monoclonal antibody
used against a tumor.
Pali- + -vi- + -zu- + -mab
humanized mab used against a virus
(RSV)
42. Types of Monoclonal Antibody
Naked Monoclonal Antibody: those without any drug
or radioactive material attached to them
Mark the cells for the immune system
Attach to receptors – block binding of growth factors
E.g.1. Trastuzumab - For advanced breast cancer (HER-2)
2. Rituximab - For B cell non Hodgkin lymphoma (CD 20)
3. Cetuximab - For advanced colorectal cancer ( HER-1)
4. Bevacizumab - For metastatic colorectal cancer (VEGF)
5. Alemtuzumab - For B cell chronic lymphocytic
leukemia. (CD 52)
43. Types of Monoclonal antibodies
Conjugated/loaded/labeled Mabs : Coupled
with drugs / toxins / radioatiactive atoms
Chemo-labeled antibodies:
○ MAbs conjugated with chemotherapeutic agents
e.g. brentuximab vedotin and ado-trastuzumab
emtansine.
○ Brentuximab vedotin, attached to a chemo drug
(MMAE) targets the CD30 antigen (present on T
and B-cells) in treatment of Hodgkin lymphoma
and non-responding anaplastic large cell
lymphoma.
○ Ado-trastuzumab emtansine, attached to DM1
chemo drug, targets the HER2 protein antigen
used for curing advanced breast cancer patients.
44. Types of Monoclonal antibodies
Immune-toxins: conjugated with toxins e.g.
Denileukin diftitox
○ used to treat some cancers (cutaneous T-cell
lymphoma and many others)
○ consists of IL-2 protein attached to a toxin (derived
from the germ causing diphtheria).
○ IL-2 normally attaches to cells that express the
CD25 antigen and thus helps in delivering the
toxin to these cells.
Radio-immune Abs: e.g. Ibritumomab,
○ an MAb against the CD20 antigen on B cells (and
lymphomas),
○ conjugated to either the radioactive isotope
indium-111 (111In) or yttrium-90 (90Y) for
treatment of lymphoma patients
45. Pharmacokinetics: mAbs
Routes of administration:
Subcutaneously (Rituximab, Trastuzumab, Adalimumab)
Intramuscularly (Palivizumab)
Intravenously
IV route: preferred because of 100% bioavailability
Route for elimination of antibodies
Via uptake & catabolism by reticuloendothelial system &
target tissue.
46. P/K: mAbs
Half-life
Chimeric : 4 –15 days
Humanized: 3 - 24 days
Recombinant human: 11– 24 days
Human antimouse antibody (HAMA) response
develops 7–10 days following exposure to murine
antibody
47. Adverse Effect of Mabs
Related to three mechanism:
Xenogenetic nature of Mab used
Suppression of physiological function
Activation of inflammatory cells or mediators after
binding of Mab to its target
48. Adverse Effect of Mabs
Naked Mabs :
Mild ; often allergic reaction on 1st infusion
Cytokine release syndrome
infusion toxicity, cytopenias
Conjugated Mabs:
More A/E’s ; depend on substances attached
Antilymphocytes Mabs
Immunosupression
increase risk of infection
Cancer
Anti TNF-α Mab
reactivation of tuberculosis
lymphomas
49. Mechanisms of Action
1. Blocking action of molecular targets
Can work antagonistically by binding a receptor to
prevent activation
Can also bind the antigen and prevent activation
2. “Magic Bullet”
Compound with target specificity is coupled with
various effector groups
○ Toxins, radionuclei, enzymes, DNA
3. Signal molecules
Coupled to mediators of apoptosis, cell division,
etc.
50. Pharmaceutical Antibodies
The fastest growing segment of the biopharmaceutical market
$65 billion in sales for 2012
there are 5 antibody drugs listed in the top 10 best selling drugs of
2012
The antibody drugs featured in the top 10 in the year 2012:
#8 = Roche’s Avastin (bevacizumab), with sales of US$6.260 billion
#7 = Johnson & Johnson’s Remicade (infliximab), with sales of
US$6.139 billion
#5 = Roche’s Hereceptin (trastuzumab), with sales of US$6.397 billion
#3 = Roche’s Rituxan (rituximab), with sales of US$7.285 billion
#1 = AbbVie’s Humira (adalimumab), with sales of US$9.265 billion.
therapeutic antibody drugs are in high demand and are playing a
significant role in the generation of revenue for drug companies
Many of the leading pharmaceutical companies have entered the mAb
sector, attracted by quicker and less costly development, higher success
rates, & premium pricing
55. Cancer
Strategies for the Mabs in Cancer:
Immune reaction directed destruction of
cancer cells
Interference with the growth & differentiation
of malignant cells
Antigen epitope directed transport of
anticancer agents to malignant cells
Anti idiotype vaccines
Variety of agents conjugated to Mabs for
selective delivery to cancer cells
56. Applications of mABs
Diagnostic tools in research and
laboratory
Different technologies in which MAbs are
used include Western blot, immunodot blot,
ELISA, radioimmuno assay (RIA), flow
cytometry, immunohistochemistry,
fluorescence microscopy, electron
microscopy, confocal microscopy
57. Applications of mABs
Diagnostic applications
MAb is used to detect pregnancy as early as a
week or two after conception by reacting with
human chorionic gonadotrophin
HIV diagnostic kits
Rapid diagnosis of hepatitis, influenza, herpes,
streptococcal & chlamydia infections
Identification & characterization of tumor specific
antigen: classification of cancer
Imaging: Localization of cancer
○ Arcitumomab : colorectal carcinoma
○ Nofetumomab : small cell lung cancer
58. Future Applications
Fight against bioterrorism
Inhalational anthrax (potent biological
terrorism) is caused by breathing the
bacterial spores of Bacillus anthracis
Raxibacumab injection is used to treat
infectious inhalational anthrax when
alternative therapies have failed
60. Limitations of MAbs
The typical doses of MAb drugs needed
for treatment are significantly higher
than those required for other drugs (or
products).
The huge demand to increase
production of these drugs and the drive
to lower the cost of these expensive
medicines is a continuous challenge to
the present industry.
61. Interesting Variations
Small antibody fragments (Fv or Fab) are also effective in
blocking cytokines
Benefit: More readily penetrate tissue
Coupling of antibody fragments to form dimers and
tetramers
Increases avidity and cross-linking
Engineered Diabodies (Bispecific mABs)
Two different antigen specificities
○ One against the target
○ The other against effectors
Can cross-link effector cells
62. Nanobodies
A Nanobody/single-domain antibody is a
peptide chain of about 110 amino acids
long, comprising one variable domain
(VH) of a heavy-chain antibody, or of a
common IgG
1989 - Raymond Hamers
Discovered in camels
Completely lack the light chain!
Same antigen affinity as their four-
chain counterparts
Structure makes them more
resistant to heat and pH
May lead to development of oral
nanobody pills
Orally available single-domain
antibodies against E. coli-induced
diarrhoea in piglets have been
developed and successfully tested
Other diseases of the gastrointestinal
tract, such as inflammatory bowel
disease and colon cancer, are also
possible targets for orally available
sdAbs
63. TO SUMMARISE :
MAbs are highly specific Abs produced by a clone of
single hybrid cells formed by fusion of B cell with the
tumor cell.
The hybridoma formed yields higher amount of
MAbs.
MAbs can be produced in vitro and in vivo .
Animals are utilized to produce MAbs, but these
antibodies are associated with immunogenicity and
ethical problems.
Recombinant DNA technology, genetic engineering
and transgenic animals are used to produce
humanized MAbs or pure human MAbs, with fewer
ADRs
Used for treatment of cancer, autoimmune disorders,
graft rejections, infections, asthma etc.
64. Finally, the dreams of Paul Ehrlich
who considered antibodies
a magic bullets have become
reality.
Monoclonal antibodies have
established themselves as the most
important and rapidly expanding
class of drugs in oncology.
Paul Ehrlich
(1854-1915)