Students of medical and allied subjects must be exposed to the concept of monoclonal antibodies for the efficient practice of clinical and laboratory medicine.
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.
Students of medical and allied subjects must be exposed to the concept of monoclonal antibodies for the efficient practice of clinical and laboratory medicine.
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.
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,
SYNTHETIC PEPTIDE VACCINES AND RECOMBINANT ANTIGEN VACCINED.R. Chandravanshi
What is a Vaccine?
A vaccine is a substance that is introduced into the body to prevent infection or to control disease due to a certain pathogen (a disease-causing organism, such as a virus, bacteria or parasite). The vaccine “teaches” the body how to defend itself against the pathogen by creating an immune response.
1 Unlike traditional pharmaceuticals, vaccines are biologics since they are made from living organisms (biological sources).
2 Specifically, vaccines are preparations of components derived from (or related to) a pathogen; they can typically induce a protective effect through one to three very small doses, in the range of micrograms to milligrams.
3 Immunity lasts for an extended period, from one year up to lifetime protection, including prevention of disease and/or related sequelae.
Synthetic peptide vaccines represent fragments of protein antigen sequences, synthesizing specific B cell and T cell epitopes offer the potential to induce diseases neutralizing immuno response with completely synthetic structure. Now it is well established that short chain peptides can be used to mimic antigenic sites of viruses and thus can be used the basics for vaccines and development. therefore, attempts have been made to synthesize such peptides which act as the serrogate immuunogens, as an alternative to the existing conventional vaccines.
Developing vaccines against infectious and epidemic diseases with the aid of Bioinformatics is now possible, by predicting epitopes on an antigen and finding possible targets for the antibody to bind. A new era of vaccine production is just ahead of us.
Watch out the ppt to know more!!!
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,
SYNTHETIC PEPTIDE VACCINES AND RECOMBINANT ANTIGEN VACCINED.R. Chandravanshi
What is a Vaccine?
A vaccine is a substance that is introduced into the body to prevent infection or to control disease due to a certain pathogen (a disease-causing organism, such as a virus, bacteria or parasite). The vaccine “teaches” the body how to defend itself against the pathogen by creating an immune response.
1 Unlike traditional pharmaceuticals, vaccines are biologics since they are made from living organisms (biological sources).
2 Specifically, vaccines are preparations of components derived from (or related to) a pathogen; they can typically induce a protective effect through one to three very small doses, in the range of micrograms to milligrams.
3 Immunity lasts for an extended period, from one year up to lifetime protection, including prevention of disease and/or related sequelae.
Synthetic peptide vaccines represent fragments of protein antigen sequences, synthesizing specific B cell and T cell epitopes offer the potential to induce diseases neutralizing immuno response with completely synthetic structure. Now it is well established that short chain peptides can be used to mimic antigenic sites of viruses and thus can be used the basics for vaccines and development. therefore, attempts have been made to synthesize such peptides which act as the serrogate immuunogens, as an alternative to the existing conventional vaccines.
Developing vaccines against infectious and epidemic diseases with the aid of Bioinformatics is now possible, by predicting epitopes on an antigen and finding possible targets for the antibody to bind. A new era of vaccine production is just ahead of us.
Watch out the ppt to know more!!!
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.
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
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.
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.
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
- 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
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.
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.
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
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.
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. What are antibodies?
3
• An antibody is a protein used by immune system to identify and
neutralize foreign objects like bacteria and viruses. Each antibody
recognizes a specific antigen unique to its target.
• The high specificity of antibodies makes them an excellent tool for
detecting and quantifying a broad array of targets, from drugs to serum
proteins to microorganisms.
• With in vitro assays, antibodies can be used to precipitate soluble antigens,
agglutinate (clump) cells, opsonize and kill bacteria with the assistance of
complement, and neutralize drugs, toxins, and viruses.
4. An antibody binds to a specific region on an antigen called an epitope.A singleantigen
can have multiple epitopes for different, specific antibodies.
4
5. Monoclonal antibodies
5
• Monoclonal antibodies are identical immunoglobulins, generated from a
single B-cell clone. These antibodies recognize unique epitopes, or binding
sites, on a single antigen. Derivation from a single B-cell clones and
subsequent targeting of a single epitope is what differentiates monoclonal
antibodies from polyclonal antibodies.
• Polyclonal antibodies are antibodies that are derived from different cell
lines.They differ in amino acidsequences.
Link: https://youtu.be/5AXApBbj1ps
6. Characters of monoclonal Antibodies
6
• Monoclonal antibodies (mAB) are 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.
• Hybridoma: A hybrid cell used as the basis for the production of antibodies
in large amounts for diagnostic or therapeutic use. Hybridomas are
produced by injecting a specific antigen into a mouse, collecting an
antibody-producing cell from the mouse's spleen, and fusing it with
a tumor cell called a myeloma cell. The hybridoma cells multiply indefinitely
in the laboratory and can be used to produce a specific antibody indefinitely.
• Myeloma cells are abnormal plasma cells (a type of white blood cell) that
build up in the bone marrow and form tumors in many bones of the body.
7. Polyclonal antibodies Monoclonal antibodies
Produced by: Many B cell clones A single B cell clone
Binds to: Multible epitopes of all
antigen used in the
immunization
A single epitope of asingle
antigen
Antibody class: A mixture of differentAb
classes (isotypes)
All of a single Abclass
Ag-binding sites: A mixture of Abs with
different antigen-binding
sites
All Abs have the sameantigen
binding sites
Potential for cross-
reactivity:
High Low
7
Differences between polyclonal and Monoclonal antibodies
8. Advantages of using MonoclonalAntibodies:
8
• Though expensive, monoclonal antibodies are cheaper to develop than
conventional drugs because it is based on tested technology.
• Side effects can be treated and reduced by using mice-human hybrid cells
or by using fractions of antibodies.
• They bind to specific diseased or damaged cells needing treatment.
• They treat a wide range of conditions.
9. Disadvantages of using MonoclonalAntibodies:
9
• Time consuming project - needs between 6 -9 months.
• Very expensive and needs considerable effort to produce them.
• Small peptide and fragment antigens may not be good antigens-
monoclonal antibody may not recognize the original antigen.
• Hybridoma culture may be subject to contamination.
• System is only well developed for limited animal and not for other
animals.
• More than 99% of the cells do not survive during the fusion process –
reducing the range of useful antibodies that can be produced against an
antigen
• It is possibility of generating immunogenicity.
11. Preparation of MonoclonalAntibodies
11
• Monoclonal Antibody production or mAb is produced by cell lines or clones
obtained from the immunized animals with the substances to be studied.
Cell lines are produced by fusing B cells from the immunized animal with
myeloma cells.
• To produce the desired mAB, the cells must be grown in either of two ways:
by injection into the peritoneal cavity of a suitably prepared mouse (in
vivo method) or by in vitro tissue culture.
13. Practical steps for production
13
Immunization of mice & isolation of splenocytes - Mice are immunized with an
antigen and later their blood is screened for antibody production. The antibody-
producing splenocytes are then isolated for in vitro hybridoma production.
Preparation of myeloma cells - Myeloma cells are immortalized cells that, once
fused with spleen cells, can result in hybridoma capable of unlimited growth.
Myeloma cells are prepared for fusion.
Fusion - Myeloma cells and isolated splenocytes are fused together to form
hybridomas in the presence of polyehthylene glycol(PEG), which causes cell
membranes to fuse.
Clone screening and picking - clones are screened and selected on the basis of
antigen specificity and immunoglobulin class.
Functional characterization - Confirm, validate and characterize (e.g. ELISA) each
potentially high-producing colony.
Scale up and wean - Scale up clones producing desired antibodies
Expansion - Expand clones producing desired antibodies (e.g. bioreactors or large
flasks).
16. 1a. Biochemical analysis
16
• Routinely used in radioimmunoassay (RIA) and enzyme-linked immunosorbent
assays (ELISA) in the laboratory.
• These assays measure the circulating concentrations of hormones (insulin,
human chorionic gonadotropin, growth hormone, progesterone, thyroxine,
triiodothyronine, thyroid stimulating hormone) and several other tissue and cell
products (blood group antigens, blood clotting factors, interferon’s, interleukins,
tumor markers).
Eg. Pregnancy by detecting the urinary levels of human chorionic gonadotropin.
Hormonal disorders analysis of thyroxine, triiodothyronine.
Cancers estimation of plasma carcinoembryonic antigen in colorectal cancer, and
prostate specific antigen for prostate cancer
17. 1b. Diagnostic imaging
17
• Radiolabeled—MAbs are used in the diagnostic imaging of diseases, and
this technique is referred to as immunoscintigraphy. The radioisotopes
commonly used for labeling MAb are iodine—131 and technetium—99. The
MAb tagged with radioisotope are injected intravenously into the patients.
• These MAbs localize at specific sites (say a tumor) which can be detected by
imaging the radioactivity. In recent years, single photon emission computed
tomography (SPECT) cameras are used to give a more sensitive three
dimensional appearance of the spots localized by radiolabeled— MAbs.
• Myocardial infarction, DVT, atherosclorosis etc.
18. 2a. Direct use of MAbs as therapeutic agents
18
• In destroying disease-causing organisms: MAbs promote efficient
opsonization of pathogenic organisms (by coating with antibody) and
enhance phagocytosis.
• In the immunosuppression of organ transplantation: In the normal
medical practice, immunosuppressive drugs such as cyclosporin and
prednisone are administered to overcome the rejection of organ
transplantation. In recent years, MAbs specific to T-lymphocyte surface
antigens are being used for this purpose
19. • In the treatment of cancer:
MAbs, against the antigens on the surface of cancer cells, are useful for the
treatment of cancer. The antibodies bind to the cancer cells and destroy
them via different pathways.
19
20. • In the treatment of AIDS: Genetic engineers have been successful to attach
Fc portion of mouse monoclonal antibody to human CD4 molecule. This
complex has high affinity to bind to membrane glycoprotein gp120 of virus
infected cells. The Fc fragment induces cell-mediated destruction of HIV
infected cells.
20
21. 2a. MAbs as targeting agents.
• The drugs can be coupled with MAb (directed against a
cell surface antigen of the cells, say a tumor) and
specifically targeted to reach the site of action.
Eg. Alkaline phosphatase for the conversion of
phosphate pro-drugs.
Carboxy peptidase for converting inactive carboxyl
pro-drugs to active drugs.
Lactamase for hydrolyzing β-lactam ring containing
antibiotics.
21
22. • MAbs in the dissolution of blood
clots:
Fibrin is the major constituent of blood
clot which gets dissolved by plasmin.
22
Plasmin in
activation
turn is formed by the
of plasminogen by
plasminogen activator.
Tissue plasminogen activator (tPA) can
be used as a therapeutic agent to
remove the blood clots.
23. • Drug delivery through liposomes coupled to tissue-specific MAbs:
Liposomes are sacs or vesicles formed spontaneously when certain lipid
molecules are exposed to aqueous environment.
Drug entrapped in liposomes that are coated with MAbs directed against
tissue-specific antigens are being tried for drug delivery.
Unfortunately, the progress in this approach has been limited, since such
liposomes do not reach the target cells.
They are retained mostly in the liver and spleen (reticuloendothelial cells),
and degraded.
23
24. 3. Protein Purification
24
• MAbs columns can be prepared by coupling them to cyanogen bromide activated
Sepharose (chromatographic matrix). The immobilized MAbs in this manner are
very useful for the purification of proteins by immunoaffinity method.
• There are certain advantages of using MAbs for protein purification. These include
the specificity of the MAb to bind to the desired protein, very efficient elution from
the chromatographic column and high degree of purification.
27. • About 75 monoclonal antibodies are currently approved by the FDA for use
in humans for treating various diseases and conditions including: cancer,
chronic inflammatory diseases, transplantation, infectious diseases and
cardiovascular diseases.
• Glenmark, which is seeking permission from MHRA, U.K. for conducting
Phase I clinical studies for one of its mAb candidate, GBR 900 mAb targeting
TrkA, the receptor of nerve growth factor to tackle chronic pain.
• India is a fertile land for mAb market due to Large patient base, growing
economy, abundant manpower and low R&D cost.
27
28. Few Commercially available mAb approved by FDA
28
https://en.wikipedia.org/wiki/List_of_therapeutic_monoclonal_antibodies
Name Trade name Target Use
Abciximab ReoPro CD41 (integrin alpha-IIb) Platelet aggregation Inhibitor
Adalimubab Humira TNF-alpha Rheumatoid arthritis, Crohn’s
Disease, Plaque psoriaris,
psoriaticArthritis
Alirocumab Praluent PCSK9 Hypercholesterolemia
Avilumab Bavencio PD-L1 Cancer
Benralizumab Facenra CD125 Asthma
Daclizumab Zenapax CD25 Organ transplant rejection
Daratumubab Darzalex CD-38 Multiple Myeloma
29. References:
29
• Shivanand P. (2010). “Hybridoma technology for production of monoclonal antibodies”
InternationalJournal of PharmaceuticalSciences Review and Research vol.1, issue 2(017)
• U. Marx et al. (1997)“MonoclonalAntibody Production” The Report and Recommendations of
ECVAMWorkshop ATLA 25,121.137,
• Edward A. Greenfield, (2014) “Antibodies:A Laboratory Manual”Cold Spring Harbor Laboratory
Press, 2nd ed. Chapter7
• Justin K.H. Liu, (2014) “The history of monoclonal antibody development Progress, remaining
challenges and future innovations” Annals of Medicine andSurgery (2014)113-116
• Andrew S., OtaviaC., (2014) “Monoclonal antibodies for the therapy of cancer” Simpson and
Caballero BMC Proceedings 2014, 8 (Suppl 4)
• WHO Guidelines on evaluation of monoclonal antibodies as similar biotherapeutic products,
2016
• BishwjitGhosal, Research project on “Study of Monoclonal Market and it’s potential in India”,
NIPER
30. • “Guidelines on evaluation of monoclonal antibodies as similar biotherapeutic
products”, WHO, 1st March,2016
• “Guideline on development, production, characterisation and specification for monoclonal
antibodies and related products”, EMA, 21st July,2016
• Biunayki R.D., “GMPCompliance for the Production of the MonoclonalAntibodyCB.Hep-1
Used as Biological Reagent | IVT Network Institute ofValidationTechnology, 2005
• https://www.thermofisher.com/in/en/home/life-science/antibodies/antibodies-learning-
center/antibodies-resource-library/antibody-methods/immunoglobulin-structure-
classes.html
• http://www.ivtnetwork.com/article/gmp-compliance-production-monoclonal-antibody-
cbhep-1-used-biological-reagent
• https://www.chromnews.com/achieving-fast-accurate-evaluation-monoclonal-antibody-
concentrations-mabs-2/
30