1.
MONOCLONAL ANTIBODIES
By:- NEHA GHANGHORIYA
M.Sc(BIOCHEMISTRY)
BU BHOPAL, MP

2.
TOPICS
COVERED:-
 1. Antibody Binding Site.
 2. Monoclonal Antibody History and Meaning.
 3. Characteristics of Monoclonal Antibody.
 4. Mechanism of mAbs generation.
 5. Diagrammatic presentation of Production of mAbs.
 6. Clinical uses of mAbs.
 7. Plasmacytoma vs B cell.
 8. Applications of mAbs.

3.
Antibody
Bindingsite

4.
Monoclonal
Antibody:-History
AndMeaning.
 Kohler and Milstein 1975.
 Fusion of Normal , Activated B cell and
Plasmacytoma(cancerous plasma cells).
 To attain high specificity, all of the antibodies must
bind With high affinity to single epitope. This high
specificity is provided By monoclonal antibodies (mAbs)

5.
CHARACTERISTICS
OFMONOCLONAL
ANTIBODIES
 Expensive Production.
 Long Production Time.
 Large Quantities of Specific Antibodies.
 Recognize a single epitope on an Antigen.
 Production is Continuous and Uniform Once a
Hybridoma is made.

6.
Mechanismof
Monoclonal
Antibodies
Generation:-
 Monoclonal antibodies are produced in vitro Using
tissue culture techniques.
 mAbs are produced by immunizing an animal, often a
mouse, multiple times with a Specific antigen.
 B cells from the spleen of the immunized animal are
then removed. Since normal B cells unable to
proliferate forever.
 They are fused with immortal, cancerous B cells called
myeloma cells, to yield Hybridoma cells.
 Then all the cells are placed in a HAT medium that
allows only hybridomas to grow as Denovo pathway is
blocked by this medium and unfused myeloma and B
cells are not able to grow and die off.

7.
mAbs
generation:-
 The Hybridomas ,which are capable of growing
continuously in culture while Producing Antibodies,
are then screened For the desired mAb.
 Those producing the desired mAb are Grown in tissue
culture.
 The culture medium is harvested periodically And
mAbs are purified from the Medium.

8.
DiagrammaticRepresentationof
mAbsProduction:-

9.
mAbsGeneration:-

10.
Clinicalusesof
mAbs:-
 Most common method to produce mAbs is mouse cells.
 Humanized mAbs:-
 Problem:-Mouse antibodies Can not be injected repeatedly into humans
, because the immune system will recognize them as being foreign and
will respond To them with neutralizing antibodies.
 Solution:-This problem can be minimized by genetically engineering
the antibody In the mouse B cell. The variable regions Of the mouse
light and heavy chain Genes are ligated to human constant regions
,And the chirmeric gene is then transferred into a host cell.
 This allows the production of mAb That is mostly “human” With only
the antigen binding site Being of mouse origin.

11.
Clinicaluseof
mAb:-
 t have been Successfully used to treat cancer with
minimal side effects.
 Example:-Humanized mAb drug Herceptin has been
helpful For the treatment of some types of breast cancer.
 Promising Technology for inexpensive mAbs:-
 The use of genetically engineered plants for production of
antibodies (plantibodies)
 This technology transforms plant cell into antibodies
factory rather than relying on the tissue culture cells,
which are expensive and technically demanding.
 Using mAbs to combate Ebola during 2014-2015 Ebola
outbreak in west Africa.A few Ebola infected patients were
treated with Zmapp, a drug Made of three antibodies.

12.
Applicationsof
mAbs:-
 Diagnosis.
 Therapeutics .
 mAbs provide higher specificity than polyclonal
antisera because they bind to a single epitope and has
high affinity.

13.
Reference:-
 Internet source.
 Kuby immunology 7th Edition, 2013.

14.
Thank you