The document discusses monoclonal antibodies (Mabs) and abzymes. It begins with an introduction to Mabs, noting they are produced by a single clone of cells and are therefore identical. It then covers the history of Mab development, characteristics of Mabs such as being derived from a single B cell clone, classification into murine, chimeric and humanized types, advantages like homogeneity and applications including diagnostic tests and cancer treatment. Problems with Mab therapy like the human immune system viewing mouse Mabs as foreign are also mentioned. The document then introduces abzymes as monoclonal antibodies with catalytic activity, discussing their sources, types and production before providing examples of reactions catalyzed by abzymes and potential applications.
DNA Protein interaction occur when a protein binds a molecule of DNA, often to regulate the biological function of DNA, usually the expression of a gene. DNA Protein interactions play very vital roles in any living cell. It controls various cellular processes which are very essential for living beings, viz. replication, transcription, recombination, DNA repair etc. There are several types of proteins found in a cell.Direct recognition occurs when the amino acid side chains of a protein interact with specific DNA bases.
Most protein-DNA interactions are mediated by direct physical interaction (hydrogen bonding or hydrophobic interactions) between the protein and the DNA base pairs.
DNA-binding proteins can be identified by many experimental techniques such as chromatin immunoprecipitation on microarrays, X-ray crystallography and nuclear magnetic resonance (NMR).
this will be useful to understand about the new topics such as abzymes, ribozymes and also isoenzymes. You have to clear that ribozymes are not protein. because all enzymes are proteins but all proteins are not enzymes except ribozymes
This is technique used widely for protein separation from a mixture and is very easy and less costly method. Slides cover all essential points about EMSA and it is quite interesting to know that how it detect and separate different proteins and their mobility shift assay.
An antibody (Ab), also known as an immunoglobulin (Ig), is a large, Y-shaped protein produced mainly by plasma cells that is used by the immune system to neutralize pathogens such as pathogenic bacteria and viruses.
Monoclonal antibodies are important reagents used in biomedical research, in diagnosis of diseases, and in treatment of such diseases as infections and cancer.
These antibodies are produced by cell lines or clones obtained from animals that have been immunized with the substance that is the subject of study.
DNA Protein interaction occur when a protein binds a molecule of DNA, often to regulate the biological function of DNA, usually the expression of a gene. DNA Protein interactions play very vital roles in any living cell. It controls various cellular processes which are very essential for living beings, viz. replication, transcription, recombination, DNA repair etc. There are several types of proteins found in a cell.Direct recognition occurs when the amino acid side chains of a protein interact with specific DNA bases.
Most protein-DNA interactions are mediated by direct physical interaction (hydrogen bonding or hydrophobic interactions) between the protein and the DNA base pairs.
DNA-binding proteins can be identified by many experimental techniques such as chromatin immunoprecipitation on microarrays, X-ray crystallography and nuclear magnetic resonance (NMR).
this will be useful to understand about the new topics such as abzymes, ribozymes and also isoenzymes. You have to clear that ribozymes are not protein. because all enzymes are proteins but all proteins are not enzymes except ribozymes
This is technique used widely for protein separation from a mixture and is very easy and less costly method. Slides cover all essential points about EMSA and it is quite interesting to know that how it detect and separate different proteins and their mobility shift assay.
An antibody (Ab), also known as an immunoglobulin (Ig), is a large, Y-shaped protein produced mainly by plasma cells that is used by the immune system to neutralize pathogens such as pathogenic bacteria and viruses.
Monoclonal antibodies are important reagents used in biomedical research, in diagnosis of diseases, and in treatment of such diseases as infections and cancer.
These antibodies are produced by cell lines or clones obtained from animals that have been immunized with the substance that is the subject of study.
It includes general introduction to antibodies; Monoclonal antibodies; comparison between Polyclonal & Monoclonal antibodies; Hybridoma Technology & Hyridoma Selection; advantages & disadvantages of mABs; Applications of mABs; Recombinant Monoclonal antibodies production through Antibody Engineering.
What are Antibody
Monoclonal Antibody (mAb)
Structure of mAb
Types of Monoclonal Antibody (mAb)
Preparation of Monoclonal Antibody
Hybridoma Technique, Phage display Technique
Application of Monoclonal Antibody
Advantage and Disadvantage of Monoclonal Antibody
Immune system; Antigens and antibodies; Epitopes, haptens and adjuvants; Complement system; Raising of anti-sera; Antigen-antibody reactions; Cellular antigens; ABO blood group system: A, B and H antigens; Bombay blood group; Rh blood group system; Blood transfusion; Transfusion reaction; Maternal reaction; Other blood groups: MN, Lewis, Lutheran, Kidd, Duffy and P systems; Secretor phenomenon; Origin testing.
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4. INTRODUCTION TO MONOCLONAL ANTIBODIES :
“MONOCLONAL ANTIBODIES are
produced by a single clone of cells. Monoclonal
antibody is therefore a single pure type of
antibody”
OR
“MONOCLONAL ANTIBODIES are
antibodies
that are identical because they were produced
by one type of immune cells.”
5. HISTORY OF MONOCLONAL ANTIBODIES
DEVELOPMENT
1890, VON BEHRING and KITASATO discovered antibodies in the
serum.
1900, EHRLICH proposed the
“ side –chain theory”.
1955, JERNE proposed
“natural selection theory”.
1964,little field a way to isolate hybrid cells.
1975 , KOHLER and MILSTEIN discovered monoclonal antibodies
first .
6. COMPARISON BETWEEN MONOCLONAL AND
POLYCLONAL ANTIBODIES
MONOLONAL POLYCLONAL
• Derived from a single B-cell
clone.
• No batch to batch variation.
• Powerful tools for clinical
diagnostic tests.
• Derived from different B
Lymphocytes cell lines .
• Batch to batch variation.
• Not powerful tools for clinical
diagnostic tests .
7. CHARACTERISTICS OF MONOCLONAL ANTIBODIES
Derived from a single B-cell clone
Each of them recognizes a specific antigen.
No batch to batch variation .
Role in immunoassay system.
Class of antibodies with identical offspring of hybridoma.
8. PRODUCTION OF MONOCLONAL ANTIBODIES
Following procedure use for their production
Inject a mouse with specific antigen.
Extract mouse spleen cells.
Extract mouse tumor cells.
Culture these cells in lab.
Mix spleen and tumor cells.
Add polyethylene glycol .
Allow the cells to grow.
Extract the hybridoma cell ,
and culture then
separately .
Culturing of cells making desired
monoclonal antibodies.
10. CLASSIFICATION OF MONOCLONAL ANTIBODIES:
3 types of Mab
• Rodent Mabs with excellent
affinities
• Also Excellent specifities
1:MURINE
SOURCE Mab
• Causes human anti chimeric
antibodies response
• e.g : Rituximab
2:CHIMERIC
MONOCLONAL
ANTIBODIES
• Contains CDRs(Clusters of
differentiation receptor) of the
rodent region e.g : Bevacizumab
3:HUMANIZED
MONOCLONAL
ANTIBODIES
12. ADVANTAGES OF MONOCLONAL ANTIBODIES :
some of their advantages are following
• It represent a single antibody
molecule.
HOMOGENITY
• Single hybridoma reacts with
same antigen epitope.SPECIFITY
IMMUNIZING
ANTIGEN
ANTIBODY
PRODUCTION
• Unlimited quantities of single
well -defined mono specific
reagent
• Not needed to produce large
quantities of antibody .
13. DISADVANTAES OF MONOCLONAL ANTIBODIES :
some of their disadvantages are following :
• Lower then polyclonal antibodies.AFFINITY
• Do not produce desired biologic
response.
EFFECTOR
FUNCTION
TIME AND
EFFORT
COMMITMENT
CROSS
REACTIONS
• Sometimes display cross
reactions
•Very large
14. APPLICATIONS OF MONOCLONAL ANTIBODIES :
Some applications of monoclonal antibodies are
following :
• To detect the presence of
substance.
Diagnostic tests
• Use for cancer and HIV
treatment .
Therapeutic
treatment
Drug
manufacturing
Organ Transplant
• Use to prevent acute rejection of
organs .
•Use in drugs to suppress immune
system.
15. PROBLEM WITH MONOCLONAL THERAPY:
Mouse antibodies are seen by Human immune system as foreign. So
then
Human anti-mouse antibodies (HAMA) produced .
Elimination of therapeutic antibodies .
Formation of Immune complexes .
16. ABZYMES
Contents
(Nazish Hanif)
• Introduction to Abzymes
• History
• Abzymes & Enzymes
(Sehar Bano)
• Sources of Abzymes
• Types of Abzymes
• Production of Abzymes
(Usama Ahmed)
• Examples of Abzymes
• Reactions catalysed by Abzymes
• Application
17. Abzymes
Definition
An abzyme (from
antibody and enzyme),
often called catalytic
antibody, is a monoclonal
antibody with catalytic
activity….
structure
18. HISTORY
The possibility of catalyzing a reaction by means of an antibody which
binds the transition state was first suggested by William P. Jencks in
1969.
In 1994, Peter G. Schultz and Richard A. Lerner received the
prestigious Wolf Prize in Chemistry for developing catalytic antibodies
for many significant reactions and popularizing their study into a
significant sub-field of enzymology
19. Abzymes VS Enzymes
Basic difference between antibodies
and enzymes is that the antibodies
binds the complementary structure in
its ground state , while enzymes bind
in high energy state
20. Sources of Abzymes
Abzymes are usually artificial constructs.
They also obtained from human and animal serum.
Found in normal humans and ii patients with autoimmune diseases.
These are capable of hydrolyzing proteins, DNA, RNA, polysaccharides
etc
21. Types Of Abzymes
PROTABZYMES
Natural abzymes with proteolytic activity are called Protabzymes
.e.g.: hydrolysis of specific proteins in patients with autoimmune
diseases such as bronchial Asthma ,multiple sclerosis.
DNA Abzymes
DNA hydrolyzing activity are called DNA abzymes.
The pathogenic role of DNA abzymes is not quite clear. However
they act as a powerful regulator of apoptosis.
22. Production of abzymes
I. Antibody molecules are produced by the immune system to
bind and neutralize foreign substances called antigens
II. Foreign proteins of bacteria and viruses called haptens , act as
antigens .
III. Transition state analogs are molecules which are more stable
than the transition state itself , but they mimic its 3D
structure .
IV. If injected into the blood stream of an animal , transition state
analogs act as haptens and elicit antibody production.
V. Abs are isolated from the serum of the animal and used as
abzymes .
VI. The oretically ,if the Ab binds to a transition state molecule, it
may be expected to catalyze a corresponding chemical
reaction by forcing substrates into transition state geometry.
24. Examples for abzymes
Hydrolysis of hydroxy ester by abzymes
Hydroxy ester Cyclic intermediate δ-lactone phenol
Anti –cyclic intermediate antibody (Abzymes)
Cyclic phosphonate ester (antigen) ,
mimic cyclic intermediate
25. Reactions catalyzed by Abzymes
1. Amide hydrolysis
2. photo cleavage
3. Photodimerization
4. Decarboxylation
5. Oxidation
6. Cyclization
7. Reduction of diketone
8. Hydrolysis of enol ethers
26. Applications
1. Synthesis of simple organic molecules
2. Drug development
1. Treat Cancer i-e:
i. Treat allergy
ii. treat viral and bacterial infection