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.
Students of medical and allied subjects must be exposed to the concept of monoclonal antibodies for the efficient practice of clinical and laboratory medicine.
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
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.
Students of medical and allied subjects must be exposed to the concept of monoclonal antibodies for the efficient practice of clinical and laboratory medicine.
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
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.
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.
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
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
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.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
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
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
Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
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.
263778731218 Abortion Clinic /Pills In Harare ,sisternakatoto
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Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
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
NVBDCP.pptx Nation vector borne disease control programSapna Thakur
NVBDCP was launched in 2003-2004 . Vector-Borne Disease: Disease that results from an infection transmitted to humans and other animals by blood-feeding arthropods, such as mosquitoes, ticks, and fleas. Examples of vector-borne diseases include Dengue fever, West Nile Virus, Lyme disease, and malaria.
NVBDCP.pptx Nation vector borne disease control program
MC 313 PPT Lecture 3 (1).pptx
1. • Mechanisms of action of antibiotics
• Structure and function of some antibiotics
• Drug resistance mechanisms
Antifungal and antiviral drugs
Industrial drug production methods
Production of penicillin/chlorotetracyclin
Use of recombinant technology in drug production
Strategy for production of human insulin by E.coli or B. subtilis
The human genomic DNA library
Transgenic animal technology
Vaccine production
PHARMACEUTICAL MICROBIAL BIOTECHNOLOGY
2. PRODUCTION OF PENICILLIN
Substrate: excess glucose leads to low yield, so
either supply slow steady input of glucose, or use
disaccharide lactose along with limiting N to enhance
drug production (stimulates production of 2° products
in idiophase).
The medium is supplemented with phenylacetic acid,
which to add benzyl side chain to final product.
The broth is removed from culture by filtration; crude
drug is extracted by adsorption, precipitation,
refinement by crystallization.
3. The kinetics of the penicillin
fermentation with Penicillium
chrysogenum.
4. Penicillin fermentation without addition of side-chain
precursors produces the natural penicillins (penicillin G)
To produce the most useful penicillins, those with
activity against gram-negative Bacteria, a combined
fermentation and chemical approach is used that leads to
the production of semi-synthetic penicillins.
Penicillin G can be chemically and or enzymatically
modified to make a variety of penicillin derivatives with
slightly different properties
These semi-synthetic penicillin derivative include
penicillin V, penicillin O, ampicillin and amoxycillin
PRODN. OF PENICILLIN AND ITS DERIVATIVES
6. PRODUCTION OF CHLORTETRACYCLINE
Chlortetracycline is produced from Streptomyces
aureofaciens.
Corn steep liquor and sucrose are used as carbon
source in large-scale production (glucose is avoided
because it causes catabolite repression of antibiotic
production).
The production scheme for chlortetracycline is shown
in the next slide.
7.
8. GEN. APPROACH FOR DESIGNING THE PRODN. OF
SYNTHETIC DRUGS
Assumption: Drugs are small molecules which function by
interacting with specific enzymes/receptor.
Goal: To develop a small molecule that specifically interacts with
active site of target enzyme/receptor.
Strategies:
- Identify target protein and determine its 3-D structure.
- Hypothesize complementary small molecules.
- Chemically synthesize the proposed molecules.
- Test the biochemical function of synthetic molecules.
- Test the physiological function of synthetic molecules
- Modify molecules to optimize efficacy.
9. USE OF RECOMBINANT TECHNOLOGY IN DRUG
PRODUCTION
Introduction
This technology takes advantage of the fact that MOs conduct
secondary metabolism leading to production of drugs.
By growing large cultures in industrial setting, one can get the
MOs to produce the crude material from which the purified
product can be obtained.
It is possible to manipulate MOs in culture and in particular,
manipulating high amounts of desired product to be produced.
Recombinant technologies take advantage of industrial
microbiology by manufacturing microbes that can produce drugs
that are not naturally synthesized by microbes i.e, bacteria or
fungi that produce non-native metabolites.
10. USE OF RECOMBINANT TECHNOLOGY IN DRUG
PRODUCTION
Several diseases are caused by a deficiency in production
of a protein or hormone.
Examples: Insulin deficiency leads to Diabetes; Factor VIII
deficiency causes hemophilia, etc.
Traditionally, the medical approach has been to isolate
these components from other animals or cadavers, when
they are similar enough to also be active in humans (e.g.
porcine insulin from pancreas tissue, somatostatin from
human cadaver brain tissue, etc).
Problem: How might it be possible to produce a MO that can
synthesize a human protein like insulin? This would be an
advantage because it would open up the possibility for large scale
11. USE OF RECOMBINANT TECHNOLOGY IN DRUG
PRODUCTION
Advantages of MOs in recombinant technology
Microorganisms are much cheaper, direct, unlimited source of
human drug products.
Example: 2 Liters of E coli can produce as much somatostatin as
that harvested from 100 sheep brains or cadavers!
12. THE APPLICATION OF GENETIC MANUPULATION TECHNIQUES
Defn. Genetic manipulation is the transfer of DNA between
different species using both in vivo and in vitro techniques, i.e.
genetic material derived from one species may be incorporated
into another species where it is expressed.
In vivo techniques: They make use of phage particles which pick
up genetic information from chromosomes of one species and
infect another species thus introducing the genetic information
from the first host. The information from the first host may then
be expressed in the second host.
In vitro techniques: It involves insertion of information into the
vector by in vitro manipulation followed by insertion of carrier
and its associated ‘extra’ DNA into the recipient cell.
13. BASIC REQUIREMENTS FOR THE IN VITRO TRANSFER AND EXPRESSION
OF FOREIGN DNA IN A HOST BACTERIUM
A ‘vector’ DNA (*plasmid or **bacteriophage ) molecule
capable self incoparation into a host cell.
Method of splicing (joining/uniting) foreign genetic information
into the vector. Use of restriction endonuclease and ligase
enzymes.
Introduction of recombinant DNA into the host cell and selecting
for their presence. Use of drug resistance markers
Assessment of the ‘foreign’ gene’s product from the population.
Achieved by testing the host cell for selected characteristics e.g.
testing if the host can confer drug resistance, etc.
*Plasmid is an extrachromosomal genetic element (DNA) found
among various strains of E. coli and other bacteria.
**Bacteriophage is the virus that infect bacterial cells.
14. BASIC REQUIREMENTS FOR THE IN VITRO TRANSFER AND
EXPRESSION OF FOREIGN DNA IN A HOST BACTERIUM
15. RESTRICTION ENZYME RECOGNITION SEQUENCE AND ENZYME
ACTION
Restriction enzymes show
specificity for certain
substrates (DNA).
They bind and digest DNA
within specific sequence of
nucleotides (restriction site
or recognition sequence)
They are commonly referred
to as 4-base pair cutters or
6-base pair cutters ie
recognize the restriction
sites with a sequence of 4 or
6 nucleotides.
Digestion of DNA by
enzyme EcoRI or KpnI
produces DNA fragments
with cohesive ends.
16. THE ERA OF GENETIC ENGINEERING/RECOMBINANT DNA
TECHNOL. IN DRUG PRODN.
Introduction
Because of the almost universal nature of the genetic
language (genetic information is encoded in linear
sequences of A, T, G, and C in a DNA strand), it is
possible to transfer human genes into bacterial cells, then
those cells should be able to synthesize human gene
products.
In practice, genes can be easily shuttled into microbeby
the use of vectors such as plasmids or viruses.
The technology takes advantage of natural routes for
genetic transmission. The bacteria will automatically
secrete the newly synthesized products.
17. 1. Isolate human insulin gene
2. Subclone into plasmid vector
3. Transform E coli with recombined plasmid
4. Select transformants that contain plasmid (antibiotic
resistance)
5. Screen transformants for ones producing the human
protein
6. Grow up large amount of the insulin-producing strain,
harvest insulin from growth medium – secreted as a
secondary metabolite.
7. Purify, test quality (absence of bacterial toxins).
STRATEGY FOR PRODUCTION OF HUMAN INSULIN BY E.
COLI OR B. SUBTILIS
18. STRATEGY FOR PRODUCTION OF HUMAN INSULIN BY E.
COLI OR B. SUBTILIS (CONT’D)
NB:
This is called a recombinant protein because it is a human protein
that is synthesized in a non-human species. In this case, an
eukaryotic protein is synthesized by a prokaryotic organism!
In some cases, the protein product is not correctly processed, so
after purification, it must be chemically modified to activate it.
Currently, S. cerevisiae (eukaryotic ) is also used in the production
of human proteins like insulin. These yeasts are also able to be
transformed by similar means to secrete desired products.
19. HUMAN GENOMIC DNA LIBRARY
• Chromosomal DNA from the
human tissue (e.g. pancreas for
insulin) is isolated and then
digested with restriction enzymes.
• A plasmid is digested with the
same enzyme and DNA ligase is
used to fuse the genonic DNA
pieces and the plasmid DNA
randomly.
• Recombinant plasmids are then
used to transform bacteria (E.
Coli), and each E. coli clone will
contain a plasmid with a human
genomic DNA fragment.
• Each clone is considered a “book”
in this “library” of DNA
fragments.
20.
21. TRANSGENIC ANIMAL TECHNOLOGY
Some current examples
• Human monoclonal antibodies (MAbs) produced by transgenic
mouse.
• Human MAbs produced by corn & soybean.
• Human Hb synthesized in transgenic pigs a blood substitute.
• Human tissue plasminogen activator (anticoagulant) produced in
transgenic goats.
• With progress in cloning technologies ( e.g. dolly the sheep), it
may soon be possible to produce herds of transgenic cattle, all
producing a human protein of interest.
• Human protein production in cows: If the gene is modified so
that it is only expressed in mammary tissue, then potentially,
human product could be isolated simply from milk.
22. MONOCLONAL ANTIBODIES (MABS)
Monoclonal antibody= An antibody produced from a
single clone of cells. It has uniform structure and
specificity.
Some terminologies
B lymphocite: a cell of the immune system that
differentiates into antibody-producing cell.
Myeloma cells: Malignant tumor of antibody-producing
cells (that no longer produces antibody of its own).
Hybridoma: a fusion of an immortal (tumor) cell with a
lymphocyte to produce an immortal lymphocyte.
23. HOW TO USE HYBRIDOMA TECHN. TO PRODUCE
MONOCLONAL ANTIBODIES
1. A mouse is immunized with the antigen of interest and left for
weeks to produce B lymphocytes.
2. The spleen tissue (rich in B lymphocytes) is removed and fused
with myeloma cells to make hybridomas.
3. The hybidomas are grown in in vitro culture containing
hypoxanthine, aminopterin and thymidine compounds ( so-
called HAT medium).
4. Fused hybrids are selected for antibody production.
5. Positive antibody-producing cells are cloned.
6. Desired clones are cultured and frozen.
7. Monoclonal antibodies are purified.
Nb. Hybridoma tumors can be kept alive in mouse
24.
25. MONOCLONAL ANTIBODIES (CONT’D)
DIFFERENCES BTW MONOCLONAL AND POLYCLONAL ANTIBODIES
Monoclonal antibodies Polyclonal antibodies
Contains single antibody
recognizing only a single
determinant on an antigen.
Contains many antibodies
recognizing many determinants
on an antigen.
Single class of antibody is
produced.
Various classes of antibodies are
produced.
Can make a specific antibody
using an impure antigen.
Can make a specific antibody
using only a highly purified
antigen.
Highly reproducible Reproducibility and
standardization is difficult.
26. VACCINE PRODUCTION
Vaccine is a biological molecule/preparation that
provides active acquired immunity to a
particular disease
Earliest strategies relied on:
Production of weakened or killed form of the pathogen -
but still able to elicit an immune response ( e.g. polio
virus)
Availability of a non-virulent strain or similar non-
pathogenic strain that could be used to elicit a cross-
reactive immune response ( e.g. cowpox, smallpox).
Limitation of this strategy: non-virulent strains have not
been identified for most infectious viruses and bacteria,
and attenuation procedures are not always 100%
complete, leading to the possibility of introducing live
pathogenic organisms via vaccine delivery itself.
27. NEW APPROACHES FOR PRODUCING VACCINE
1. Creating recombinant viral particles to serve as vaccine
vectors e.g. vaccinia virus.
Attenuated forms of vaccinia virus (which is responsible
for cowpox and smallpox) have existed for many years.
This is an enveloped, DNA virus similar to many other
animal viruses (enveloped instead of naked like many
bacteriophages).
It is possible to grow this virus in culture: grow up
mammalian cell line, then infect it with vaccinia virus,
culture produces large amounts of viral particles, which
bud from the surface of the cultured cells.
28. NEW APPROACHES FOR PRODUCING VACCINE
(CONT’D)
2. Overexpressed viral proteins or combinations of viral
proteins in bacteria or yeast are isolated and used as
immunogen in vaccine mixtures.
Isolated proteins may not present the same traits as in
the native viral particles.
It is possible to genetically modify the viral genome so
that non-vaccinia proteins are produced.
Using genetic recombination it is possible to produce
protein antigens for use in synthesizing vaccines. (see
diagram below)
Imunogen Antigen that is capable of inducing immune
response.
29. CLONING OF FOREIGN DNA INTO VACCINIA VIRUS
• Gene for different viral
surface protein can be
inserted and will then be
expressed on the surface of
infected cells and ultimately
incorporated into the
envelope of the new viral
particles.
• Modified viral particles are
harvested and used, as
vaccine – essentially these
particles present the new viral
antigen to the immune system
so that it can respond with a
primary response.
• Gen. Modified vaccinia is
used as vaccine: it is safer,
more reproducible and can be
30. OTHER VACCINES UNDER INVESTIGATION/ DEVELOPMENT
Hepatitis B – recombinant protein used as immunogen.
Malaria vaccine – Plasmodium falciparum surface
protein expressed in recombinant yeast cultures, trial
conducted in the Gambia.
HIV vaccine –1st attempts were made in early 1990s
using recombinant gp120 protein produced in
mammalian cells. The trials proved ineffective.
AIDS vaccine (VAX BB) trials - currently underway in
US and Thailand – includes a mixture of different
gp120 variants.
HIV1 env/gag/pol genes - inserted into vaccinia viral
genome under vaccinia viral gene control.
gp120/CD4 complex- under lab investigation as
potential immunogen vaccine.