The document discusses hybridoma technology, which involves fusing antibody-producing B cells from an immunized animal with myeloma cells to produce a hybrid cell line that secretes monoclonal antibodies specific to a single epitope. This overcomes limitations of earlier polyclonal antibody approaches. Key points covered include the history of the technology's development, the cell fusion and selection process, advantages like unlimited production of highly specific antibodies, and applications such as diagnostic assays.
Gene editing in veterinary vaccine development: Status of the scienceILRI
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Gene editing in veterinary vaccine development: Status of the scienceILRI
Presentation by Lucilla Steinaa at a CGIAR webinar on 'Genome Editing in Agriculture: Innovations for Sustainable Production and Food Systems', 6 October 2020.
In recent years, antibodies have become increasingly accepted as therapeutics for human diseases, particularly for cancer, viral infection and autoimmune disorders.
Monoclonal antibodies (Mabs) have been used as diagnostic and analytical reagents since hybridoma technology was invented in 1975.
“man-made antibodies.” was named by Cesar Milstein, who was one of the inventors of monoclonal antibody technology.
Until the late 1980’s, antibody technology relied primarily on animal immunization and the expression of engineered antibodies.
Baculoviruses are the most important tool for the study of basic virology and biology. Baculoviruses are highly beneficial viruses since they do not infect man or plants but they do provide effective natural biological control of many insect species. They are also versatile vectors for the expression of proteins for basic research and medical applications.
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.
In recent years, antibodies have become increasingly accepted as therapeutics for human diseases, particularly for cancer, viral infection and autoimmune disorders.
Monoclonal antibodies (Mabs) have been used as diagnostic and analytical reagents since hybridoma technology was invented in 1975.
“man-made antibodies.” was named by Cesar Milstein, who was one of the inventors of monoclonal antibody technology.
Until the late 1980’s, antibody technology relied primarily on animal immunization and the expression of engineered antibodies.
Baculoviruses are the most important tool for the study of basic virology and biology. Baculoviruses are highly beneficial viruses since they do not infect man or plants but they do provide effective natural biological control of many insect species. They are also versatile vectors for the expression of proteins for basic research and medical applications.
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.
BIOTECHNOLOGY IS
CHALLENGING SUBJECT TO TEACH AND UNDERSTAND ......
ITS A VERY INTERESTING TO LEARN ABOUT HYBRIDOMA TECHNOLOGY .. THEIR PRODUCTION AND
APPLICATION ALSO ....
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Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
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June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
Hybridoma cell lines Introduction, Production, Growth, Limitations and Techniques
1.
2. T A B L E O F C O N T E N
T S
Introductio
n
01
History
02
Growth
medium
03
Production
04
Techniques and
examples
05
Advantages /
Disadvantages
06
Application
s
07
3. A hybridoma is a cell line arising from one hybrid
cell that is capable of secreting a monoclonal
antibody specific to one epitope of your antigen
permanently in culture.
The hybrid cell is produced through the fusion of
specific antibody producing B-cell from an
immunized animal (usually a mouse, rat or rabbit)
and which has a finite lifespan, with a cell from an
“immortal” cultured myeloma cell line
I N T R O D U C T I O
N
4. H I S T O R Y
196
4
Way developed
to isolate hybrid
cells using HAT
media
197
5
Georges J.F Kholer
and Caser Milstein
provide the proof
Hybridoma
Technology
1984
Awarded
Nobel prize
5. 2
1
BEFORE HYBRIDOMA CELL-
LINES
The
problem is
that
antibody-
producing
cells
isolated
from an
organism
die in a
short time
Only produce
polyclonal
antibodies, did
not immortalize
the antibody-
producing cells,
so it required
repeated animal
use to obtain
more antibodies
6. G R O W T H M E D I U M
01
Bicarbonate
uffer, with L-
Glutamine
02
Good quality
Foetal Bovine
Serum
03
HEPES buffer,
without L-
Glutamine
05
PEG
(Polyethylene
Glycol)
04
HAT
(Hypoxathine,
Aminopterin,
Thymidine)
7. P R O C E D U R E
1
2
3
4
5
6
IMMUNIZATION
SELECTION&INJECTI
ON
SCREENING
ANTIBODY
PRODUCTION
PREPARATION
MYELOMA CELLS
FUSION
MYELOMA&SAPLEE
N CELLS IN PEG
HYBRIDOMA
CELLS
PROLIFERATE&GRO
W
SELECTION
mAbs DESIRED
SPECIFICITY
8. P R O C E D U R E
7
8
CLONING
BY LIMITING
DILUTION IN
CULTURE MEDIA ISOLATION
EXPAND OR FREEZ
9.
10. idiotype
(antigenic
specificity).
• B-CELL CANCER
• MEMBRANE BOUNDED
ANTIBODIES HAVE
SAME IDIOTYPE
mAb specific for
idiotype
• Anti-idiotype antibodies
produce in mouse
• Make humanized in
bacteria
• Inject into human
• Bind on idiotype and
activate complement
system
64-year-old
man
• TERMINAL B-
LYMPHOMA
• Metastasized
to the liver,
spleen, bone
marrow, and
peripheral
blood.
11.
12. A D V A N T A G E S / B E N E F I
T S
Generation of highly
specific mAbs
Specific to a single epitope of the
target antigen
Host’s natural ability
Hybridoma generation process takes
advantage of a host animal's natural
ability to generate functional, highly
specific and high-affinity mAbs.
Advantages over classic
hybridization
Transgenic mouse can produce a
variety of specific antibodies, from
which candidates can be isolated, and
it is likely that these will be less
immunogenic than the chimeric
antibodies
No species level barriers
Shown success in isolation of
hybridoma across phylogenetically
distinct species and led to the
isolation of mAbs against human
targets, conserved in the rodent
Robust & immortal source
Basic practical advantage: once the
hybridoma clones are established,
the production of mAb becomes
simple and efficient
Hybridoma serves as an immortal
source of monoclonal antibody.
Highly reproducible and scalable,
unlimited production source.
Employs different species
Initially, limited to murine antigens –
with advancement –a vast range of
antigens and from different species
like rabbits, humans, chickens, goats,
sheep, cows, mice, guinea pigs, and
rats.
Indigenous nature
It is the most favored method due to
its indigenous nature to preserve
natural cognate antibody pairing
information and preserves innate
functions of immune cells.
Use of chicken egg yolk
For antibody production represents a
reduction in animal use (ethical
issues) as chicken produces a larger
amount of antibodies than laboratory
rodents.
In liquid nitrogen for several years
making them virtually immortal for
continuous mAb production for a
long period.
Cryopreservation
13. Time
Consumin
g
tedious process - takes 6 months to
year time.
Risk of
Contaminatio
n.
Hybridoma culture may be
subject to contamination.
Expensiv
e
Hybridomas are sometimes
difficult and expensive to
maintain in culture.
Effort
Requiring
needs considerable effort to
produce them.
Requirement
of Purified
Ag
immunization of animals with these purified
recombinant proteins in formulation with
adjuvants - to alteration of native
conformation of these proteins -undesired
immune response in animals
D I S A D V A N T A G E S / L I M I T A T I
O N S
