Nanotechnology and Targeted drug delivery systems

2. Monoclonal antibodies are the
antibodies produced in the laboratory by a clone
of cultured cells that make one specific
antibody.
An Antibody is a protein used by the
immune system to identify and neutralize the
foreign objects like bacteria and virus. Each
antibody recognizes a specific antigen unique to
its target.

3.  Derived from different B-Lymphocytes
cell lines.
 These antibodies are non-specific and
heterogeneous in nature.
 They have the ability to bind with
different antigens.
 Batch to batch variation affecting
antibody reactivity. Derived from a single B-Cell clone.
 These antibodies are specific and
homogeneous in nature.
 They have ability to bind with single
antigen.
 No batch to batch variation & not
affecting antibody reactivity.

5. AdvantagesAdvantages
Homogeneity:
Monoclonal antibody represents a single
antibody molecule that binds to antigens with the
same affinity and promote the same effectors'
functions.
Specificity:
The product of a single hybridoma reacts
with the same epitope on antigens.
Immunizing antigen:
Need not be pure or characterized and is
ultimately not needed to produce large quantities
of antibody.

6. Cont….
Selection:
It is possible to select for specific epitope
specificities and generate antibodies against a
wider range of antigenic determinants.
Antibody production:
Unlimited quantities of a single well defined
monospecific reagent.

7. Affinity:
Average affinity of monoclonal antibodies are
generally lower than polyclonal antibodies when
they bind with antigens.
Effectors' functions:
The monoclonal antibody may not produce the
desired biologic response compared to
polyclonal antibodies.
Specificity:
Monoclonal antibodies against conformational
epitopes on native proteins may loss reactivity

8. Cont….
Cross-reactions:
Monoclonal antibodies sometimes may
display unexpected cross-reactions with unrelated
antigens.
Time & effort commitment:
Monoclonal antibodies having very large
time and effort commitment compared to
polyclonal antibodies.

11. Conventional production of Monoclonal
antibodies
Based on fusion of murine myeloma cells
and spleen cells different cell cultures methods are
classified as follows:
1.Rollerbottle cell culture process:
Roller bottle provides conditions that
favor the transfer of oxygen and temperature
control without aeration, agitation propellers or
circulation pumps.
The bottle is mounted on a turn table
which gives homogeneity of growth and aeration of
the culture medium.

12. 2. Suspended cell culture process:
Bioreactors are commonly used for
monoclonal antibody production in submerged
mammalian cells.
Bioreactors are:-
• Stainless steel stirred tank bioreactor
• Airlift reactor
• Disposable bioreactors
Stainless steelstirredtankbioreactor:
 cleaning and sterilization protocols are
involved.
 maintain the cultivation parameters like gas
transfer coefficient, agitation, aeration,
temperature maintenance and PH
are well
controlled and regulated.

13. Disadvantage:-
It can cause cell lysis and lead to loss
in monoclonal antibody productivity (decreases
production).

14. Airlift reactors:
 The air flows through the columns length to the
top of the bioreactor as degassed culture.
 Medium flows in the opposite direction to the
bottom of the reactor.
 It creates a constant gentle mixing of the
medium as well as proper culture aeration.
 Finally formation of monoclonal antibodies.

16. Disposablebioreactor:
 The wave system consists of a sterile plastic
bag lying on a rocking platform.
 The bag is half filled with cultivation medium
and remaining half filled with a gas mix of interest.
 The platform motion creates an undulation
movement in the culture.
 It creates an efficient aeration and culture
mixing without causing shear damage.
 Production of monoclonal antibodies.

18. Othermethods:
 Membrane bound cell culture process
 Micro carrier cell culture process
These methods are also used for
production of monoclonal antibodies.

19. Monoclonal antibodies can be broadly categorized
as :
 Diagnostic application
 Therapeutic application
 Catalytic monoclonal antibodies (Abzymes)
A. Diagnostic applications:
 monoclonal antibody can be used to detect
pregnancy in 14days.
 Detection of low levels of “Human chorionic
gonadotropin” in urine and serum.
 monoclonal antibodies are used for “Western blot
test” and “Immuno dot blot test” to detect the
protein on a membrane.

20. Cont….
 Useful in “Immuno histo chemistry process” of
detecting antigens.
 Useful in “Immuno fluorescence test” to detect
the substance in a frozen tissue section/live cells.
 Monoclonal antibodies can also be used to
purify a substance with techniques called “Affinity
chromatography”.

21. B. Therapeutic application:
O rg an transplantatio n- “Orthrochrome OKT3” was
the first monoclonal antibody for the treatment of
solid organ transplant rejection.
Bo ne m arro w transplantatio n- Monoclonal
antibodies are evaluated for “Graft v/s Host
disease” in bone marrow transplantation.
Cance r tre atm e nt- Monoclonal antibodies act
directly binding to a cancer specific agents and
induces immunological response to cancer cells.
Used in Radio im m uno the rapy.
Co njug ate d m o no clo nalantibo dy the rapy- toxins
or radio active isotopes bound to the region of
monoclonal antibodies and these antibodies bind
to the tumor cells and radio activity will kill the

22. FDA approvedmonoclonal antibodies
(cancertreatment):
 Bevacizumab – Breast & lung cancer
 Cetuximab – Colon, head & neck cancer
 Panitumumab – Colon cancer
 Trastuzumab – Breast cancer
 Alemtuzumab – Chronic lymphocytic
leukemia

23. C. Catalytic monoclonalantibody:
 Binding of an antibody to antigen and enzyme to
its substrates both involves weak and non-covalent
interactions which shows “High Affinity” and “High
Specificity”
 It involves both “Antigen-Antibody interaction”
and “Enyme-Substrate interaction”
 Auto antibody fingerprinting using “Dip sticks”
can be done in “Rheumatic diseases”
 Monoclonal antibodies are used to target drugs