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TOPIC : MONOCLONAL ANTIBODIES(Mab) AND
ABZYMES
 AMNA SHOUKAT 7211
 USAMA 7225
 AMINA BASHIR 7227
 SEHAR BANO 7272
 MEHTAB REHMAN 7273
 FAIQA ZAFAR 7259 (Group Leader)
 NAZISH HANIF 7276
 SUBJECT : IMMUNOLOGY
SUBMITTED TO : DR. MUHAMMAD RIZWAN JAVED
SUBMISSION DATE : 27th,OCT ,2014
 3rd SEMESTER
 DEPARTMENT : BIOINFORMATICS AND BIOTECHNOLOGY
MONOCLONAL ANTIBODIES
Contents
(Amna Shoukat)
1: Introduction
2: History
3: Comparison b/w monoclonal antibodies
& Polyclonal antibodies
(Faiqa Zafar)
4: Characteristics
5: Production
(Amina Bashir)
6: Classification
7: Advantages
8: Disadvantages
(Mehtab Rehman)
9: Applications
10: Problem with monoclonal antibody therapy
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.”
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 .
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 .
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.
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.
PRODUCTION OF MONOCLONAL ANTIBODIES
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
TYPES OF MONOCLONAL ANTIBODIES
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 .
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
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.
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 .
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
Abzymes
Definition
 An abzyme (from
antibody and enzyme),
often called catalytic
antibody, is a monoclonal
antibody with catalytic
activity….
structure
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
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
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
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.
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.
Transition
state
Transition
state analog
(act as Ag)
Substrate
Ab complementary
to transition state
Pdt
mice
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
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
Applications
1. Synthesis of simple organic molecules
2. Drug development
1. Treat Cancer i-e:
i. Treat allergy
ii. treat viral and bacterial infection
REFERENCES
 http://www.meds.com/immunotherapy/monoclonal_
antibodies.html
 Kuby Immunology by Richard A. Goldsby, Thomas J.
Kindt and Janis Kuby.
 WWW.slideshare.net
 Enzymology –T. Devasena
• Wikipedia

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Monoclonal and abzymes

  • 1.
  • 2. TOPIC : MONOCLONAL ANTIBODIES(Mab) AND ABZYMES  AMNA SHOUKAT 7211  USAMA 7225  AMINA BASHIR 7227  SEHAR BANO 7272  MEHTAB REHMAN 7273  FAIQA ZAFAR 7259 (Group Leader)  NAZISH HANIF 7276  SUBJECT : IMMUNOLOGY SUBMITTED TO : DR. MUHAMMAD RIZWAN JAVED SUBMISSION DATE : 27th,OCT ,2014  3rd SEMESTER  DEPARTMENT : BIOINFORMATICS AND BIOTECHNOLOGY
  • 3. MONOCLONAL ANTIBODIES Contents (Amna Shoukat) 1: Introduction 2: History 3: Comparison b/w monoclonal antibodies & Polyclonal antibodies (Faiqa Zafar) 4: Characteristics 5: Production (Amina Bashir) 6: Classification 7: Advantages 8: Disadvantages (Mehtab Rehman) 9: Applications 10: Problem with monoclonal antibody therapy
  • 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
  • 11. TYPES OF 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.
  • 23. Transition state Transition state analog (act as Ag) Substrate Ab complementary to transition state Pdt mice
  • 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
  • 27. REFERENCES  http://www.meds.com/immunotherapy/monoclonal_ antibodies.html  Kuby Immunology by Richard A. Goldsby, Thomas J. Kindt and Janis Kuby.  WWW.slideshare.net  Enzymology –T. Devasena • Wikipedia