- Monoclonal antibodies are identical antibodies produced by a single clone of B cells or hybridoma. They are produced by fusing B cells from an immunized animal with myeloma cells.
- The hybridoma technique involves immunizing an animal, isolating spleen B cells, fusing them with myeloma cells using polyethylene glycol, and screening clones to identify those that produce the desired antibody.
- Hybridomas are immortal cell lines that can produce large quantities of identical monoclonal antibodies directed against a specific antigen or epitope. This technique allows mass production of antibodies for research, diagnostic, and therapeutic uses.
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.
CLONAL SELECTION THEORY IS AN SCIENTIFIC THEORY IN IMMUNOLOGY THAT EXPALINS THE FUNCTION OF CELLS OF THE IMMUNE SYSTEM IN RESPONSE TO SPECIFIC ANTIGEN INVADING THE BODY.
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.
CLONAL SELECTION THEORY IS AN SCIENTIFIC THEORY IN IMMUNOLOGY THAT EXPALINS THE FUNCTION OF CELLS OF THE IMMUNE SYSTEM IN RESPONSE TO SPECIFIC ANTIGEN INVADING THE BODY.
As opposed to common belief, the measurement of growth in cell culture is fairly simple. Most of the tecchniques that are applied for measurement of microbial growth can be applied to cell culture.Of course with some modification. This presentation exactly explains growth measurement techniques with respect to cell culture. At the end you will also find sample multiple choice questions for practice.
INTRODUCTION
ROLE IN CELL LINE CHARACTERIZATION
CAUSES OF TRANSFORMATION
METHODS OF TRANSFECTION
CHARACTERISTICS OF TRAANSFORMED CELLS
GENETIC INSTABILITY
IMMORTALIZATION
ABRERANT GROWTH CONTROL
TUMORIGENECITY
CHROMOSOMAL ABERATION
APPLICATION
CONCLUSION
REFERENCE
B Cell Receptor & Antibody Production-Dr C R MeeraMeera C R
Antibody production is the function of B lymphocytes. These slides describe the structure of B cell receptor and steps involved in antibody production by B lymphocytes
As opposed to common belief, the measurement of growth in cell culture is fairly simple. Most of the tecchniques that are applied for measurement of microbial growth can be applied to cell culture.Of course with some modification. This presentation exactly explains growth measurement techniques with respect to cell culture. At the end you will also find sample multiple choice questions for practice.
INTRODUCTION
ROLE IN CELL LINE CHARACTERIZATION
CAUSES OF TRANSFORMATION
METHODS OF TRANSFECTION
CHARACTERISTICS OF TRAANSFORMED CELLS
GENETIC INSTABILITY
IMMORTALIZATION
ABRERANT GROWTH CONTROL
TUMORIGENECITY
CHROMOSOMAL ABERATION
APPLICATION
CONCLUSION
REFERENCE
B Cell Receptor & Antibody Production-Dr C R MeeraMeera C R
Antibody production is the function of B lymphocytes. These slides describe the structure of B cell receptor and steps involved in antibody production by B lymphocytes
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
Students of medical and allied subjects must be exposed to the concept of monoclonal antibodies for the efficient practice of clinical and laboratory medicine.
BIOTECHNOLOGY IS
CHALLENGING SUBJECT TO TEACH AND UNDERSTAND ......
ITS A VERY INTERESTING TO LEARN ABOUT HYBRIDOMA TECHNOLOGY .. THEIR PRODUCTION AND
APPLICATION ALSO ....
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
3. 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 are 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.
4. History of mAB development:
1964 Littlefield developed a way to isolate hybrid cells from 2 parent cell lines using
the hypoxanthine-aminopterin-thymidine (HAT) selection media.
1975 Kohler and Milstein provided the most outstanding proof of the clonal selection
theory by fusion of normal and malignant cells
1990 Milstein produced the first monoclonal antibodies.
Paul Ehrlich at the beginning of the 20th century theorized that a cell under threat
grew additional side-chains to bind the toxin, and that these additional side chains broke
off to become the antibodies that are circulated through the body. It was these
antibodies that Ehrlich first described as "magic bullets" in search of toxins.
5. Characters of Monoclonal Antibodies:
Monoclonal antibodies (mAb) are a 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.
A hybridoma cell line is formed by the fusion of a one Bcell lymphocyte
with a myeloma cell.
Some myeloma cells synthesize single mAb antibodies naturally
6. Monoclonal Antibody Production method:
Monoclonal Antibody Production technology was developed in 1975.
Since its development it has been very important in the modern medical
science with the diagnosis, therapy, research and even basic science
today. It is still largely dependent upon animal testing however. Because
it requires immunization of mice in order for them to create the
antibodies to be grown.
Monoclonal Antibody Production or mAb is produced by cell lines or clones
obtained from the immunized animals with the substance 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 (the in vivo, or mouse ascites, method) or by in
vitro tissue culture.
The vitro tissue culture is the method used when the cells are places in
culture outside the mouse's body in a flask.
7. The previous picture is showing a mouse being immunized against a target
cell “X”. This will allow the mouse to produce antibodies for that will target
against the “X” antigen.
Once the mouse has formed antibodies to the “X” antigen the cells are then
isolated in the mouse’s spleen. Monoclonal antibodies are produced by fusing
single antibody-forming cells to tumor cells grown in culture. The resulting
cell is called a hybridoma. Hybridoma cells are continuously growing cell line
generated by the fusion of a myeloma cell and a normal cell that are capable
of producing antibodies.
Each hybridoma will produce relatively large quantities of identical antibody
molecules. Because the hybridoma is multiplying in culture, it is possible to
produce a population of cells, each is producing identical antibody
molecules. These antibodies are called "monoclonal antibodies" because
they are produced by the identical offspring of a single, cloned antibody
producing cell.
8.
9. Practical steps in monoclonal antibody production:
Immunize animal
Isolate spleen cells (containing antibody-producing B cells)
Fuse spleen cells with myeloma cells (e.g. using PEG – polyethylene glycol)
Allow unfused B cells to die
Add aminopterin to culture to kill unfused myeloma cells
Clone remaining cells (place 1 cell/well and allow each cell to grow into a clone of cells)
Screen supernatant of each clone for presence of the desired antibody
Grow the chosen clone of cells in tissue culture indefinitely.
Harvest antibody from the culture supernatant.
(If you’re a biotech company) charge about $1,000-$2,000 per mg.
11. Hybridoma creates Monoclonal Antibodies:
Monoclonal antibodies are typically made by fusing
myeloma cells with the spleen cells from a mouse that has
been immunized with the desired antigen. However, recent
advances have allowed the use of rabbit B-cells.
Hybridoma cells grown in tissue culture.
12. Monoclonal – Diagnostic Use:
A monoclonal antibody can be used to detect pregnancy
only 14 days after conception. Other monoclonal
antibodies allow rapid diagnosis of hepatitis, influenza,
herpes, streptococcal, and Chlamydia infections.
They can be used to detect for the presence and
quantity of this substance, for instance in a Western
blot test (to detect a substance in a solution) or an
immunofluorescence test.
Monoclonal antibodies can also be used to purify a
substance with techniques called immunoprecipitation
and affinity chromatography.
13. Monoclonal Antibodies for Cancer treatment:
Possible treatment for cancer involves
monoclonal antibodies that bind only to
cancer cells specific antigen and induce
immunological response on the target cancer
cell (naked antibodies). mAb can be
modificated for delivery of [toxin],
radioisotope, cytokine.
14. Possible side effects of Monoclonal Antibodies:
Some mAbs can also have other side effects that are related to the
antigens they target. For example, bevacizumab (Avastin), an mAb
that targets tumor blood vessel growth, can cause side effects such as
high blood pressure,bleeding, poor wound healing, blood clots, and
kidney damage.
Fever
Chills
Weakness
Headache
Nausea
Vomiting
Diarrhea
Low blood pressure
Rashes
16. Definition:
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 mammal) with
an antigen that provokes an immune response. A type of white blood
cell, the B cell that produces antibodies that bind to the antigen are
then harvested from the mouse. These isolated B cells are in turn
fused with immortal B cell cancer cells, a myeloma, to produce a
hybrid cell line called a hybridoma, which has both the antibody-
producing ability of the B-cell and the exaggerated longevity and
reproductivity of the myeloma.
17. In 1975, Kohler and Milstein first fused lymphocytes
to produce a cell line which was both immortal and a
producer of specific antibodies. The two scientists
were awarded the Nobel Prize for Medicine in 1984
for the development of this "hybridoma." The value of
hybridomas to the field was not truly appreciated
until about 1987, when MAbs were regularly
produced in rodents for diagnostics.
Kohler and Milstein
18.
19. Immunisation of a mouse
Isolation of B cells from the spleen
Cultivation of myeloma cells
Fusion of myeloma and B cells
Separation of cell lines
Screening of suitable cell lines
in vitro (a) or in vivo (b) multiplication
Harvesting
Method of Hybridoma technique:
20. Methodology:
A hybridoma, is produced by the injection of a specific antigen into a
mouse, procuring the antigen-specific plasma cells (antibodyproducing
cell) from the mouse's spleen and the subsequent fusion of this cell
with a cancerous immune cell called a myeloma cell.
Once spleen cells are isolated from the mammal’s spleen, the B cells
are fused with myeloma cells.
The hybrid cell, which is thus produced, can be cloned to produce
many identical daughter clones.
21. These daughter clones then secrete the immune cell product. Since these
antibodies come from only one type of cell (the hybridoma cell) they are
called monoclonal antibodies.
The advantage of this process is that it can combine the qualities of the
two different types of cells; the ability to grow continually, and to produce
large amounts of pure antibody.
22.
23. HAT Selection:
HAT medium (Hypoxanthine Aminopterin Thymidine) is used for preparation
of monoclonal antibodies.
Laboratory animals (e.g.. mice) are first exposed to an antigen to
which we are interested in isolating an antibody against.
Once spleen cells are isolated from the mammal, the B cells are fused
with immortalized myeloma cells, which lack the HGPRT
(hypoxanthine-guanine phosphoribosyl transferase) gene using
polyethylene glycol.
Fused cells are incubated in the HAT (Hypoxanthine Aminopterin
Thymidine) medium.
24. Myeloma cells die, as they cannot produce nucleotides by the de
novo or salvage medium blocks the pathway that allows for
nucleotide synthesis.
Unfused B cells die as they have a short life span.
Only the B cell-myeloma hybrids survive, since the HGPRT gene
coming from the B cells is functional. These cells produce antibodies
(a property of B cells) and are immortal (a property of myeloma
cells)