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Monoclonal Antibodies

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22 views10 pages

Monoclonal Antibodies

Uploaded by

vaishnavipawar988
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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NOVEL DRUG DELIVERY SYSTEM

MONOCLONAL ANTIBODIES
-Miss Vaishnavi Pawar
- Miss Mrunal Rokade
ANTIBODIES:
Basically Antibodies are special molecules in our body which helps
the body fight against the Infections caused by various foreign
microorganisms like Bacteria,Viruses,Toxins,and many other foreign
particles.
- These Antibodies are of different shapes and sizes and basically ‘Y’ shaped.

HOW DO ANTIBODIES WORK ?


MONOCLONAL ANTIBODIES :
Monoclonal Antibodies are artificially produced Antibodies and are
very specific for their intended targets.

-When a Immunogen for eg : a toxoid enters a body;Humoral Immune system is


provoked and thus Antibodies are produced in the organism against that specific
Immunogen at different parts or regions of the foreign substance.
-These different parts of an antigen i.e Foreign substance also called as ‘Antigenic
determinants or Epitopes’ are recognized by these specific
Antibodies and then Antibodies interact with the Antigen sites and form a antigen
antibody complex which further phagocytize or eliminate the
foreign substance.
-Each antibody is specific for a single epitope and each antibody is product of a single
B cell clone.
These Antibodies of unique apecificity for a specific epitope derved from
a single B cell are termed as ‘Monoclonal Antibodies’.
APPLICATIONS OF MONOCLONAL ANTIBODIES :
-Monoclonal Antibodies are essential tools for many molecular immunology
investigations.
- They have wide applications in detecting and identifying serum analytes,
cell markers ,and pathogenic agents due to their exquisite specificity.
- Very Helpful in Diagnosis of various Autoimmune diseases .in Diagnosis and
treatment of Cancer as they target specific cells.
Types of Monoclonal Antibodies:
a) Murine Source Monoclonal Antibodies(Mouse derived) :
-These are derived from B cells of rodents.
- They have excellent affinities and specificities and are generated using
conventional Hybridoma Techniques.
eg : Rituximab
b) Chimeric Monoclonal Antibodies :
-These Combine Mouse variable regions with human Constant regions
- Affinity and specificity remains unchanged.
eg : Cetuximab
c) Humanized Monoclonal Antibodies :
- They contain only the CDRs(Complemantarity Determining Regions)of rodent
variable regions grafted onto Human Framework regions. i.e. More Human
sequence and minimal Rodent variable regions.
eg : Trastazumab
d)Human Monoclonal Antibodies :
- These contain Fully Human Sequence derived from Human B cells.
Currently Available Techniques for Production of Human Monoclonal Antibodies
include:
1. The Hybridoma Technique: Human B lymphocytes +Myeloma cells
2. The EBV(Epstein Bar Virus) immortalization Technique
3. The EBV Hybridoma Technique :Combination of a) and b)
Production of Monoclonal Antibodies:
This Process involves 6 steps
• Immunization
• Cell fusion
• Selection of Hybridoma
• Screening of Products
• Cloning and Propagation
• Characterization and Storage
IMMUNIZATION:
- It is done by Injecting an imunogen i.e. Antigen mixed with an Adjuvant.
It is given Intradermally or Subcutaneously at multiple sites repeatedly.
-Final dose before killing the animal is given Intravenously.
-Spleen is then asceptically removed and disrupted by mechanical or
enzymatic methods to release the cells.

CELL FUSION:
- Formed Lymphocytes are fused in vitro with Myeloma cells.
-This fusion is carried out by adding PEG to the palette containing both spleen
and myeloma cells.

SELECTION OF HYBRIDOMA:
- Above mixture is cultured in HAT medium for 7-10 days.
-This medium allows growth of only hybrid cells Since,Myloma cells are unable
to synthesize HGPRT,Hence free myeloma cells die.
SCREENING OF PRODUCTS :
- Screening is done for antibody specificity.
-For this,Common tests like ELISA are carried out to detect the presence of
antigen specific antibodies.
- The hybridomas secreted are called as Monoclonal Antibodies.

CLONING AND PROPAGATION :


- The single hybrid cell producing desired antibody is isolated and cloned.
Two techniques are usually employed for this :
a) Limiting Dilution Method: Suspension of Hybridoma cells is serially diluted
until each dilution is having one hybrid cell. This ensures that the Antibody produced
is Monoclonal.
b) Soft agar Method: In this method,Hybridoma cells are grown insoft agar to
form colonies and the colonies are Monoclonal in nature.
ADVANTAGES OF MONOCLONAL ANTIBODIES :
-They are Homogenous in nature i.e. Produced from only one type of cells.
-They are specific to a particular Antigen with particular Epitope.

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