Transcript CHRONIC MYELOGENOUS LEUKAEMIA

MONOCLONAL ANTIBODIES
SEMINAR PRESENTATION
Performed by
PASCHALIS KOURELIAS
MSc BIOMEDICAL IMMUNOLOGY
EAST LONDON UNIVERSITY
STRATFORD 09 /12/2003
WELCOME ABOARD
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WELCOME TO THE MOST EXCITING
DISCOVERY OF MODERN MOLECULAR
IMMUNOLOGY.
THE JOURNEY TO THE WORLD OF
MONOCLONAL ANTIBODIES STARTS
JUST……… NOW !!!!!!!
WHAT IS AN ANTIBODY ?
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ANTIBODIES or IMMUNOGLOBULINS ARE THE
SECRETED FORM OF THE B-CELL ANTIGEN
RECEPTOR (BCR)
THEY’RE MAIN ROLE IS TO BIND TO A FOREIGN
ANTIGEN, AND THEN MEDIATE SECONDARY
EFFECTS(ACTIVATION OF COMPLIMENT
SYSTEM,RECRUITMENT OF PHAGOCYTES,
OPSONIZATION).
THEY ARE REMARKABLY DIVERSE AND PROVIDE
DIFFERENT COMBINE SITES TO RECOGNIZE THE
MILLIONS OF ANTIGENIC SHAPES IN THE
ENVIRONMENT.
WHAT IS MONOCLONAL
ANTIBODY? PART 1
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MONOCLONAL ANTIBODIES (m-Abs),
ARE Abs WHICH ARE GENERATED BY
EXPANDING AND IMMORTALIZING
SINGLE CLONES OF B-CELLS,EACH OF
WHICH HAS A DEFINED SPECIFICITY.
WHAT IS MONOCLONAL
ANTIBODY ? PART 2
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BECAUSE OF THE DEFINED SPECIFICITY
THE mAbs RECOGNIZE JUST ONE ONLY
EPITOPE ON THE ANTIGEN.
THE MONOCLONAL Abs WHICH DERIVED
FROM ONE CLONE ARE ALMOST OF A
SINGLE ISOTYPE
HOW WE CAN MAKE THEM I
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THEY PRODUCED BY CELL LINES OR
CLONES OBTAINED FROM ANIMALS
(mainly mice) THAT HAVE BEEN
IMMUNIZED WITH THE SUBSTANCE THAT
IS SUBJECT OF STUDY.
AN ALTERNATIVE WAY IS TO TRASFORM
B-CELLS (HUMAN B-CELS)BY INFECTED
THEM WITH EPSTEIN-BARR VIRUS.
HOW WE CAN MAKE THEM II
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TO PRODUCED THE DESIRED mAb,
THE CELLS MUST BE GROWN IN
EITHER OF TWO WAYS:
BY INJECTION INTO THE ABNOMINAL
CAVITY OF A SUITABLY PREPARED
MOUSE.
BY TISSUE CULTURING CELLS IN
PLASTIC FLASKS.
HOW WE CAN MAKE THEM III
HOW WE CAN MAKE THEM
SIGNIFICANCE-USE OF mAbs
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mAbs ARE USED EXTENSIVELY IN
BIOMEDICAL RESEARCH.
IN DIAGNOSIS OF A DISEASE, INFECTION.
IN TREATMENT OF A DISEASE
IN TREATMENT OF CANCER
IN SEROLOGICAL ASSAYS
AS MARKERS IN IDENTIFICATION OF
PROTEINS,CARBOHYDRATES,NUCLEIC
ACIDS.
USE OF MONOCLONAL Abs
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MONOCLONAL Abs WERE ENVISAGED AS
FORMING A VERY SPECIFIC HIGH
AFFINITY DELIVERY SYSTEM FOR A TOXIN
OR RADIOISOTOPE.
TO DELIVER A DAMAGING STIMULUS TO
THE TUMOUR CELL OR VIRALLY
INFECTED CELL WITHOUT HAVING ANY
DETRIMENTAL EFFECT ON NORMAL
NEIGHBOURING CELLS.
EFFECTS OF THE USE OF mAb
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BECAUSE OF MICE (FOREIGN ORIGEN) THEY
COULD INITIATE A XENOGENEIC EFFECT.
The production of human anti-mouse Abs
HAMA
LIMITED USEFULNESS AND APLLICATION AS
THERAPEUTIC AGENTS.
The monoclonal Abs rapidly cleared from recipient’s
system, by immune response.
HUMAN MONOCLONAL Abs
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THE PROCESS IS MUCH MORE COMPLEX IN
THE IMMUNISATION STAGES AND IN FUSION
STEP.
MORE DIFFICULT TO PRODUCE
IMMORTALISATION OF HUMAN ANTIDODY
SECRETING CELLS.
ALTERNATIVE OPTIONS BECOME NECESSARY,
AND BECOME AVAILABLE THANKS TO THE
MAJOR ADVANCES OF GENETIC ENGINEERING
AND RECOMBINANT DNA TECHNOLOGY.
CHIMERIC ANTIBODIES
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GENETIC ENGINEERING AND GENE
MANIPULATION HUMANISED THE MOUSE
MONOCLONAL Abs.
POSSIBLE TO CLONE A V- region FROM A
MOUSE ANTIBODY AND EXPRESS THIS
GENE ALONG WITH A C- region OF THE
REQUIRED TYPE.
XENOGENEIC EFFECT IS LIMITED
VAST REPERTOIRE OF V-regions OF mAbs CAN BE UTILISED.
MOSAIC V-REGIONS
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TO FURTHER HUMANISE THE MICE
MOBOCLONAL Abs, MOSAIC V-region Abs,
HAVE BEEN PRODUCED.
ONLY PORTIONS OF MOUSE ORIGIN ARE
THE CDR’s.
HUMAN FWR AND C-regions.
LESS OF THE MOLECULE IS XENOGENEIC.
CHOICE OF C-regions ALLOWS THE
TAILORINGOF THE MOLECULE TO A
PARTICULAR PURPOSE
SUMMARY
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MONOCLONAL Abs DERIVED FROM JUST ONLY A SIGLE
CLONE. ALWAYS FROM THE SAME ISOTYPE.
GENERATE FROM MICE BY FUSING SPLEEN CELLS
WITH A CANCER CELL (MYELOMA) or
BY USING IN VITRO TECHNIQUES.
HUMAN m-Abs ARE VERY LIMITED IN PRODUCTION DUE
TO SEVERAL DIFFICULTIES.
THEY HAVE A GREAT USE IN MOLECULE MEDICINE. IN
DIAGNOSIS-TREATMENT OF DISEASES INCLUDING
CANCER.
SEVERAL METHODS HAVE BEEN DEVELOPED FOR HUMANISED
THE MICE MONOCLONAL Abs.
CHIMERIC ANTIBODIES, MOSAIC V-regions,
ANY QUESTIONS ?
ANY QUESTIONS ????
REFERENCES
1.Day,NE. Varghese C. (2001).25 years of monoclonal
Antibodies. Immunology Today. Volume 32, No 8.
2.Avidan B, Sonneberg A.(2002). Role of monoclonal
antibodies in molecular medicine.
Journal of Immunology. Vol 97:1130-1135.
3. Blot W.J, Gefferson B. (2001). Humanised mice
antibodies. Current opinion in Immunology35:403409.
4. Gammon. MD.(2000).Monoclonal antibody. New
England Journal Med. 405:362-365.