This document summarizes monoclonal antibodies (mAbs). It discusses how mAbs are produced by fusing B cells from an immunized animal with myeloma cells, generating hybridomas that produce antibodies targeting a single epitope. The document outlines the history of mAbs, their characteristics including high specificity and continuous production. It also describes the clinical uses of mAbs such as treating cancer and Ebola, as well as applications in diagnosis and therapeutics due to their high specificity and affinity.

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.

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

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

16. Thank you