Hybridoma technology allows for the production of monoclonal antibodies through the fusion of antibody-producing B cells with myeloma cells. This fusion produces hybridoma cells that are selected using HAT medium. The hybridoma cells are then screened to identify those that produce the desired monoclonal antibody. Monoclonal antibodies produced through this process are highly specific and useful for applications such as disease diagnosis, purification of proteins, and cancer therapy. However, some limitations include immune responses against mouse antibodies in humans and contamination risks with hybridoma cell cultures.
PREPARATION OF BACTERIAL VACCINES:
Steps involved in killed bacterial vaccine preparation:
1. Selection of an antigen:
The exact strain or strains to be incorporated for preparation of bacterial vaccine.
Eg. Cholera vaccine: smooth strains of the two serological types Inaba and Ogawa
TABC vaccine: O and H antigens in S. typhi and S. paratyphi microorganisms and these organisms also contains Vi antigen.
Each strain is carefully checked for freedom from variation and absence of contaminating organisms.
Use of microbes in industry. Production of enzymes-General consideration-Amyl...Steffi Thomas
Industrial uses of microbes, properties of useful industrial microbes, various industrial products, production of enzymes-general consideration-amylase, catalase, peroxidase, lipase, protease, penicillinase, procedure for culturing bacteria and inoculum preparation, submerged fermentation and solid state fermentation, uses of different enzymes
BOTECHNOLOGY IS CHALLENGING SUBJECT TO TEACH AND UNDERSTAND ALSO .....THEIR INTERESTING PART IS TO LEARN ABOUT MICROBIAL BIO TRANSFORMATION WITH BIOCHEMICAL REACTIONS
genetic engineering, principles, b pharma 6th sem, biotechnology
What is a gene ?
Definition
History
Process
Molecular tools of genetic engineering
Restriction enzymes
History of restriction enzyme
Mechanism of action
Types of restriction enzymes
Application of restriction enzymes
Blunt ends
Sticky ends
transgenic
cisgenic.
knockout organism.
Host organism vector
TRANSGENIC PLANTS
DOLLY THE SHIP
TRANSGENIC ANIMALS
PREPARATION OF BACTERIAL VACCINES:
Steps involved in killed bacterial vaccine preparation:
1. Selection of an antigen:
The exact strain or strains to be incorporated for preparation of bacterial vaccine.
Eg. Cholera vaccine: smooth strains of the two serological types Inaba and Ogawa
TABC vaccine: O and H antigens in S. typhi and S. paratyphi microorganisms and these organisms also contains Vi antigen.
Each strain is carefully checked for freedom from variation and absence of contaminating organisms.
Use of microbes in industry. Production of enzymes-General consideration-Amyl...Steffi Thomas
Industrial uses of microbes, properties of useful industrial microbes, various industrial products, production of enzymes-general consideration-amylase, catalase, peroxidase, lipase, protease, penicillinase, procedure for culturing bacteria and inoculum preparation, submerged fermentation and solid state fermentation, uses of different enzymes
BOTECHNOLOGY IS CHALLENGING SUBJECT TO TEACH AND UNDERSTAND ALSO .....THEIR INTERESTING PART IS TO LEARN ABOUT MICROBIAL BIO TRANSFORMATION WITH BIOCHEMICAL REACTIONS
genetic engineering, principles, b pharma 6th sem, biotechnology
What is a gene ?
Definition
History
Process
Molecular tools of genetic engineering
Restriction enzymes
History of restriction enzyme
Mechanism of action
Types of restriction enzymes
Application of restriction enzymes
Blunt ends
Sticky ends
transgenic
cisgenic.
knockout organism.
Host organism vector
TRANSGENIC PLANTS
DOLLY THE SHIP
TRANSGENIC ANIMALS
UNIT-5 Protein Engineering: Brief introduction to protein engineering,Use of ...Shyam Bass
UNIT-5 6th Sem B.PHARMA PHARMACEUTICAL BIOTECHNOLOGY)
Protein Engineering: Brief introduction to protein engineering, Use of microbes in industry, Production of enzymes-general considerations, Amylase, Catalase, peroxidase, Lipase Basic principles of genetic engineering
BY- SHYAM BASS
UNIT 4 Microbial genetics:Transformation,Transduction,Conjugation,Plasmids an...Shyam Bass
(6th Sem B.Pharma Pharmaceutical Biotechnology)
Microbial genetics:
• Transformation,
• Transduction,
• Conjugation,
• Plasmids and transposons,
• Study of the production of - Penicillins, Citric acid, Vitamin B12, Glutamic acid,
Griseofulvin,
• Blood Products: Collection, Processing, and Storage of whole human blood,Dried
human plasma, Plasma substitutes
BY- SHYAM BASS
General method of the preparation of Bacterial Infections, Toxoids, Viral vaccine, Antitoxins, Serum-immune blood derivatives and other products relative to immunity.
UNIT 6 Fermentation technology, Fermenters, Study of Media, types of fermenta...Shyam Bass
UNIT-6 6th Sem B.Pharma Pharmaceutical Biotechnology-
Following slides include-
Fermentation technology and biotechnological products :
Fermentation methods and general requirements
Study of media
Equipment
Sterilization methods
Aeration process
Stirring
large scale production fermenter design and its various controls
BY- SHYAM BASS
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.
Pharmaceutical biotechnology ..in that different blotting techniques such as ELISA , western blotting and southern blotting.There applications and their advantage and disadvantage with their diagrams
UNIT-5 Protein Engineering: Brief introduction to protein engineering,Use of ...Shyam Bass
UNIT-5 6th Sem B.PHARMA PHARMACEUTICAL BIOTECHNOLOGY)
Protein Engineering: Brief introduction to protein engineering, Use of microbes in industry, Production of enzymes-general considerations, Amylase, Catalase, peroxidase, Lipase Basic principles of genetic engineering
BY- SHYAM BASS
UNIT 4 Microbial genetics:Transformation,Transduction,Conjugation,Plasmids an...Shyam Bass
(6th Sem B.Pharma Pharmaceutical Biotechnology)
Microbial genetics:
• Transformation,
• Transduction,
• Conjugation,
• Plasmids and transposons,
• Study of the production of - Penicillins, Citric acid, Vitamin B12, Glutamic acid,
Griseofulvin,
• Blood Products: Collection, Processing, and Storage of whole human blood,Dried
human plasma, Plasma substitutes
BY- SHYAM BASS
General method of the preparation of Bacterial Infections, Toxoids, Viral vaccine, Antitoxins, Serum-immune blood derivatives and other products relative to immunity.
UNIT 6 Fermentation technology, Fermenters, Study of Media, types of fermenta...Shyam Bass
UNIT-6 6th Sem B.Pharma Pharmaceutical Biotechnology-
Following slides include-
Fermentation technology and biotechnological products :
Fermentation methods and general requirements
Study of media
Equipment
Sterilization methods
Aeration process
Stirring
large scale production fermenter design and its various controls
BY- SHYAM BASS
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.
Pharmaceutical biotechnology ..in that different blotting techniques such as ELISA , western blotting and southern blotting.There applications and their advantage and disadvantage with their diagrams
Topics included :- Introduction to monoclonal antibody; Principle for creation of hybridoma cells and steps involved in it; Second generation monoclonal antibodies; Advantages, disadvantages and applications (Diagnostic and therapeutic) of MAbs.
Hybridoma Technology ( Production , Purification , and Application ) Sakshi Ghasle
Hybridoma technology revolutionized the field of immunology by enabling the production of monoclonal antibodies with high specificity and affinity. This presentation delves into the principles of DNA hybridoma technology, highlighting its significance in antibody production, therapeutic applications, and biomedical research. Learn about the key steps involved in generating hybridomas, from immunization to antibody screening, and discover the potential of recombinant DNA techniques in enhancing antibody engineering. Whether you're a student, researcher, or industry professional, this overview will provide valuable insights into the innovative world of hybridoma technology."
Uncover the wide-ranging applications of monoclonal antibodies in areas such as cancer therapy, autoimmune diseases, infectious diseases, and beyond. Learn about the latest advancements in antibody engineering and the development of novel therapeutic modalities, including bispecific antibodies, antibody-drug conjugates, and immune checkpoint inhibitors.
Whether you're a seasoned researcher or a newcomer to the field, this SlideShare presentation serves as a valuable resource for understanding the principles, techniques, and applications of hybridoma technology in modern biomedicine. Join a journey through the fascinating world of monoclonal antibodies and the groundbreaking science behind their creation.
Unlock the potential of hybridoma technology and propel your research to new heights. Dive into this SlideShare presentation now and explore the limitless possibilities of monoclonal antibody production with hybridoma technology.
Hybridoma
Hybridomas are cells that have been engineered to produce a desired antibody in large amounts, to produce monoclonal antibodies.
Monoclonal antibodies can be produced in specialized cells through a technique now popularly known as hybridoma technology.
Hybridoma technology was discovered in 1975 by two scientists, G. Kohler and C. Milstein, were awarded Noble prize for physiology and medicine in 1984.
Various diagnostic tools now a days relied on the Hybriodoma technology and monoclonal antibodies,so this presentation will give some basic information about mAb and Hybridoma technology.
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
A well established method to produce monoclonal antibodies specific to antigen of interest.
used in,various therapies like in tumor cell removal from bone marrow, treatment in acute renal failure , in malignant leukemic cells , in immuno purification also have serological importance.
Monoclonal Antibodies and their applicationsDr.S.Selvaraj
A specific and single type of antibody that is produced by hybridoma cells.
This technique was achieved by George Kohler and Cesar Milstein in 1975. (Nobel Prize, 1984)
Antibodies (or) immunoglobulins is a protein - produced by B- lymphocytes (Plasma cell) in mammals.
Antibodies used by immune system to identify and neutralize foreign objects like bacteria and viruses.
Each antibody recognizes a specific antigen unique to its target.
ACTIVE CONSTITUENT OF DRUGS USED IN DIABETIC THERAPYAswinisasidharan
ACTIVE CONSTITUENT OF DRUGS USED IN DIABETIC THERAPY, GYMNEMA SYLVESTRE, GYMNEMIC ACID, SALACIA RETICULATE, CHEMICAL CONSTITUENT, MECHANISM OF ACTION, USES.
Flavonoids, Definition, Classification, General Methods for the Structural determination of Flavonoids, Structural Elucidation of Quercetin, References.
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.
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
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.
Anti ulcer drugs and their Advance pharmacology ||
Anti-ulcer drugs are medications used to prevent and treat ulcers in the stomach and upper part of the small intestine (duodenal ulcers). These ulcers are often caused by an imbalance between stomach acid and the mucosal lining, which protects the stomach lining.
||Scope: Overview of various classes of anti-ulcer drugs, their mechanisms of action, indications, side effects, and clinical considerations.
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
These lecture slides, by Dr Sidra Arshad, offer a quick overview of physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
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.
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.
2. HYBRIDOMA TECHNOLOGY
• Hybridoma technology is a method of forming hybrid cell lines
(hybridomas) by fusing a specific antibody-producing B-cell
with a myeloma cell (cancerous cell).
HISTORY
• The production of monoclonal antibodies was first done by
Niels K.J. Georges, J.F. Kohler and Cesar Milstein in 1975.
3. PRINCIPLE
• It is based on the fusion between myeloma cells (malignant plasma cells)
and spleen cells from a suitably immunized animal.
• Spleen cells die in a short period under ordinary tissue culture conditions
while myeloma cells are adopted to grow permanently in culture.
• From the growing hybrids, individual clones can be chosen that secrete
the desired antibodies.
4. PROCEDURE
1. Immunization.
2. Isolation of myeloma cells.
3. Fusion between spleen cell and myeloma cell.
4. Selection of HAT medium.
5. Isolation of hybridoma cell.
6. Screening of hybridoma cell.
5. Immunization of specific animal
Antigen immunized to animal (like mice) via intravenously.
In spleen, activates B-cell in which it produces plasma cell
Plasma cell produces monoclonal antibodies
Isolation of plasma cell from spleen of animal.
6. Isolation of myeloma cells
• Myeloma cells are cancerous cells which is isolated from
bone-marrow.
• Generally immortal in nature and has multiplication
property.
7. Fusion of spleen cell and myeloma cell
• It requires PEG medium for fusion
• It can also done by electro fusion.
• Fusion between spleen cell and myeloma cell produces five
different types of cells;
● Fused plasma
● Fused myeloma
● Hybridoma
● Unfused plasma
● Unfused myeloma
9. Selection of HAT medium
• Before multiplication of Anti-body, it has to synthesize new
copy of DNA and for that it require synthesis of nucleotide.
• Synthesis of nucleotide have two pathways:
•
1. Salvage pathway
2. De-novo Synthesis
11. Isolation Of Hybridoma Cells
Only the hybridoma cells survive in HGPRT Medium.
HGPRT
● Fused plasma -present
● Fused myeloma -absent
● Hybridoma -present
● Unfused plasma -present
● Unfused myeloma -absent
Spleen cells have HGPRT Enzyme while myeloma doesn’t have it.
12. Screening of hybridoma cell
• ELISA screening method which done by incubating hybridoma
culture in which secondary enzyme gets conjugate and
formation of coloured product shows positive hybridoma.
• Hybridoma cells producing desired antibody was identified by
screening.
• Cloning, Propagation, Production & Purification has been done.
14. MERITS
• It produces highly specific antibodies in abundant amounts.
• The clones developed are far cheaper than the traditionally
• employed animals.
• It produces high quality products, to avoid the batch to batch
variation.
•
15. DEMERITS
• Many patients develop immune resistance to monoclonal
antibodies produced by mice, as these are foreign proteins.
• Hybridoma culture may be subjected to contamination.
• System is only well developed for mouse and rat, not for the
other animals.
• More than 99% of the cells do not survive during the fusion
process.
16. APPLICATIONS
1. Serological - Identification of ABO blood group.
2. Diagnosis - Early detection of Pregnancy
HIV
Cancer
3. Immunopurification - Purification of individual interferones.
Inactivation of T lymphocytes
4. Therapy - Removal of tumour cell.
Acute renal failure
Leukemia
17. References
• Current Trends in Biotechnology By Dr S satyalakshmi, pageno; 294-
302.
• A Textbook of Pharmaceutical Biotechnology By Mr Akshay Shashikant
Patil, Dr Md Rageeb Md Usman, Dr Vijay Mishra, Dr Bhushankumar S
Sathe, pageno; 126-134.
• Textbook of Biotechnology By Sathyanarayanan U, pageno; 213-226.