Dr/ Ahmed Abdellatif Assistant Professor at the Faculty of Pharmacy, - Qassim University, Kingdom of Saudi Arabia. - Al-Azhar University, Egypt.

1. Dr/ Ahmed Abdellatif
Assistant Professor at Faculty of Pharmacy,
- Qassim University, Kingdom of Saudi Arabia.
- Al-Azhar University, Egypt.

3. Any of the highly specific antibodies produced in lar
ge quantity by the clones of a single hybridoma cell.
Monoclonal antibodies (mAb or moAb)
are monospecific antibodies that are made by
identical immune cells that are all clones of a unique
parent cell. Monoclonal antibodies
have monovalent affinity, in that they bind to the
same epitope.

4.  Antibodies have important uses beyond fighting
infections in the body.
 Production of long-lasting monoclonal antibodies is a
recent invention and it is used in both medicine and
research.
 Monoclonal Antibody: a stable antibody which can be
used over a period of time

5. 1. Inject a mouse with a specific antigen to stimulate its
immune system to produce necessary antibodies.
2. Extract mouse spleen cells (containing B-
lymphocytes) and culture them in the lab.
3. Extract mouse tumor cells, which grow continuously,
and culture them in the lab.
4. Mix spleen cells and tumor cells on the same plate and
culture.

6. 5. Add polyethylene glycol – this causes some B-
lymphocytes to fuse with tumor cells to produce
a hybrid cell called a hybridoma.
6. Grow the cells under conditions that allow only
hybridoma cells to survive.
7. Extract the cells, culture them separately and test
the medium around each cell for the specific
antibody of interest.
8. Culture the cells making the desired antibody
and use as needed.

8.  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.

9.  A pregnant woman has the hormone human chorionic
gonadotrophin (HCG) in her urine.
 Monoclonal antibodies to HCG have been produced.
These have been attached to enzymes which can later
interact with a dye molecule and produce a color
change.

10.  Pregnancy test contains three regions – reaction
region, test region and control region
◦ Reaction region: contains monoclonal anti-HCG antibodies
linked to enzyme
◦ Test region: contains polyclonal anti-HCG antibodies,
which will bind to HCG molecules bound to monoclonal
anti-HCG antibodies. Also contains dye molecules which
will be activated if monoclonal antibodies bind to polyclonal
anti-HCG antibodies
◦ Control region: contains anti-mouse antibodies and dye
molecules which will be activated if monoclonal antibodies
bind here

11.  When pregnant woman’s urine travels up the
pregnancy test, HCG will bind to monoclonal
antibodies in reaction region
 Movement of the urine will move the monoclonal
antibodies up to the test region
 Monoclonal antibodies with bound HCG will bind to
antibodies in test region and activate dye molecules,
producing a color change
 Any unbound monoclonal antibodies will continue to
travel to control region and will bind to anti-mouse
antibodies, activating dye molecules and producing a
color change

13.  The test of HIV infection
is based on detecting the
presence of HIV
antibody in the patient’s
blood serum.

14. a) HIV antigen is attached to the plate.
b) Patients serum passed over the plate. Any HIV
antibody in the patients serum will be attached to the
antigen already on the plate.
c) A second antibody which is specific to the HIV
antibody is passed over the plate. This antibody will
attach to the concentrated HIV antibody on the plate.
This second antibody has an enzyme attached to its
structure.
d) Chromogen dye is passed over the complex of
concentrated HIV antibody/conjugated antibody.
e) The enzyme will turn the chromogen to a more
intense color. The more intense the color, the greater
the HIV antibody level. This would be the a positive
result for a HIV test.

15.  Cancer cells carry specific tumor-associated antigens
(TAA) on their plasma membrane.
 Monoclonal anti-TAA antibodies have been produced.
 Drugs which kill tumor cells or inhibit key proteins in
tumor cells are attached to monoclonal anti-TAA
antibodies.
 Cancer cells are specifically targeted, avoiding damage
to healthy host cells.

16.  Many patients develop the immune response to
monoclonal antibodies produced in mice, as these are
foreign proteins.
 Genetically engineered antibodies are being perfected
to avoid triggering the immune response.

17. Stem Cells

18.  A cell that has the ability to continuously divide and
differentiate (develop) into various other kind(s) of
cells/tissues

19.  ‘Blank cells’ (unspecialized)
 Capable of dividing and renewing themselves for
long periods of time (proliferation and renewal)
 Have the potential to give rise to specialized cell
types (differentiation)

20. Stem cell
type Description Examples
Totipotent
Each cell can develop into
a new individual
Cells from early (1-3
days) embryos
Pluripotent
Cells can form any (over
200) cell types
Some cells of
blastocyst (5 to 14
days)
Multipotent
Cells differentiated, but
can form a number of other
tissues
Fetal tissue, cord
blood, and adult
stem cells

21. This cell
Can form the
Embryo and placenta
This cell
Can just form the
embryo
Fully mature

23.  Multipotent stem cells
– limited in what the
cells can become

24. Embryonic Stem Cells
Mainly from IVF

26. Why do researchers study embryonic
stem cells?
 Tissue-specific stem cells are limited in their
differentiation potential (blood  blood)
 Stem cells from some tissues are inaccessible
 Some tissue-specific stem cells don’t self-renew well
 Some tissues may not have stem cells!

27. blastocyst Blastocyst inner mass cells
8-cell stagecleavage

29.  Skin
 Fat Cells
 Bone marrow
 Brain
 Many other organs & tissues
Adult Stem Cells
An undifferentiated cells found among
specialized or differentiated cells in a tissue
or organ after birth

30. Induced Pluripotent Stem Cells

31. Stem Cell Applications
• Tissue repair
- nerve, heart, muscle, organ, skin
• Cancers
• Autoimmune diseases
- diabetes, rheumatoid arthritis, MS

32. Tissue Repair
• Regenerate spinal cord, heart tissue
or any other major tissue in the body.

33. Heart Disease
• Adult bone marrow stem cells injected
into the hearts are believed to improve
cardiac function in victims of heart
failure or heart attack

34. Leukemia and Cancer
• Studies show leukemia patients treated
with stem cells emerge free of disease.
• Injections of stem cells have also
reduces pancreatic cancers in some
patients.
Proliferation of white cells

35. Rheumatoid Arthritis
• Adult Stem Cells may be helpful in
jumpstarting repair of eroded cartilage.

36. Type I Diabetes
• Pancreatic cells do not produce insulin
• Embryonic Stems Cells might be
trained to become pancreatic islets cells
needed to secrete insulin.

37. • Differentiation of stem cells into
mature, functional cells
• Potential for tumor formation
• Immune rejection

38.  Embryonic Stem cells are derived from extra
blastocysts that would otherwise be discarded
following IVF.
 Extracting stem cells destroys the developing
blastocyst (embryo).
 Questions for Consideration-
 Is an embryo a person?
 Is it morally acceptable to use embryos for
research?
 When do we become “human beings?”

39. Thank You