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1
MONOCLONAL ANTIBODIES
PRESENTED BY: SRIJA BOSE, SRIDEBESH GHORUI
ROLL NO.:18601916033,18601916034
REGISTRATION NO.: 161860210106,161860210105
B.PHARM, 3RD YEAR,6TH SEMESTER,SECTION-A
GURU NANAK INSTITUTE OF PHARMACEUTICAL SCIENCE AND TECHNOLOGY
222
 Introduction
 Principle for the creation of hybridoma cells
 Production of monoclonal antibody
 Principles of HAT selection
 Applications of monoclonal antibodies
 Latest news on monoclonal antibodies
 Conclusions
 References
3
INTRODUCTION
Monoclonal antibody (MAb) is a single type of
antibody that is directed against a specific
antigenic determinant(epitope).
MAbs are immunoglobulins or a fragment of an
immunoglobin produced by a single clone of cells.
They can be obtained from immortalized B-
lymphocytes that are cloned and expanded as
continuous cell lines(murine and human
monoclonal antibodies) or from rDNA engineered
mammalian or bacterial cell lines(engineered
monoclonal antibodies).
When a foreign body or antigen molecule enters
into the body, the immune response begins.
4
PRODUCTION OF MONOCLONALANTIBODY
• The establishment of hybridomas and production of MAbs involves the following steps:
1. Immunization
2. Cell fusion
3. Selection of hybridomas
4. Screening the products
5. Cloning and propagation
6. Characterization and storage
55
PRODUCTION OF MONOCLONALANTIBODY
6
PRINCIPLE FOR THE CREATION OF HYBRIDOMA CELLS
• Hybridoma cells can be produced by the fusion of an
antibody producing cell and a myeloma cell.
• The mammalian cells can synthesize nucleotides by two
pathways:-
A)De novo synthesis
B)Salvage pathway
77
PRINCIPLES OF HAT SELECTION
1. When the major synthetic pathways are blocked by the folic acid analogue aminopterin, the cell should
employ the salvage pathway. This pathway essentially contains the enzyme HGPRT.
2. HGPRT- cells die in the presence of HAT because both the major synthetic pathway and the salvage
pathway are blocked virtually.
3. Fusion of the HGPRT- myeloma cells with the HGPRT+ spleen cells allows growth in HAT by
incorporating the missing enzyme for the salvage pathway.
88
HAT SELECTION
99
APPLICATIONS OF MONOCLONALANTIBODIES
• Cancer therapy or immunotoxicology
• Bone marrow transplantation
• Organ transplantation
• Protein purification
• Disease diagnosis
• Gene cloning and expression
1010
LATEST NEWS ON MONOCLONALANTIBODIES
• MAb pembrolizumab prolongs the survival in patients with squamous cell
carcinoma.(Review on JAN 18,2019)
• Tychan has successfully conducted the Phase 1 trial of TY014, a first-in-class monoclonal
antibody candidate treatment for yellow fever. (13th DEC 2018)
• The meta-analysis of 10 randomized controlled trials has confirmed that human MAb
denosumab can more effectively increase the bone mass in patients with low bone mineral
density or osteoporosis. Any other biphosphate therapy can not do. (13th DEC 2018)
• Sarlumab developed in collaboration with Sanofi and Regeneron Pharmaceuticals, is a MAb
targeting interleukin 6 receptors approved by the FDA. (May 2017)
• Roche’s Emicizumab, the last approved MAb drug in 2017, targets the coagulation factors IXa
and X, thereby promoting the clotting process.
1111
CONCLUSIONS
• Pathogens can be detected in a small number of hours sparing few days of
culturing of cells.
• Since for the human health care, the subject of production and use of antibodies
has become a very important area of research, not only for the academic purpose
but also for its relevance to industrial growth for diagnosis and therapeutics, MAb
can be utilized for it.
• MAb technology in conjunction with recombinant DNA technology has
successfully led to the reconstruction of chimeric, humanized and fully human
antibodies.
1212
REFERENCES
• ‘Sambamurthy K., Kar Ashutosh’, “Pharmaceutical Biotechnology” Reprint of First Edition:
2008, New Age International (P) Ltd. Publishers, ISBN:81-224-1911-9, Chapter 1: Immunology
and Immunological Preparations, Subunit 5: Antigen Antibodies Reactions and Their
Applications (5.1.5 Monoclonal Antibodies), Page 53-58.
• Dr. U Satyanarayana, “Biotechnology” Reprint Of First Edition:2010, Books And Allied (P)
Ltd., ISBN:81-87134-909, Chapter17 Monoclonal Antibody (Hybridoma Technology) Page-
213-214.
1313
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Monoclonal antibodies

  • 1. 1 MONOCLONAL ANTIBODIES PRESENTED BY: SRIJA BOSE, SRIDEBESH GHORUI ROLL NO.:18601916033,18601916034 REGISTRATION NO.: 161860210106,161860210105 B.PHARM, 3RD YEAR,6TH SEMESTER,SECTION-A GURU NANAK INSTITUTE OF PHARMACEUTICAL SCIENCE AND TECHNOLOGY
  • 2. 222  Introduction  Principle for the creation of hybridoma cells  Production of monoclonal antibody  Principles of HAT selection  Applications of monoclonal antibodies  Latest news on monoclonal antibodies  Conclusions  References
  • 3. 3 INTRODUCTION Monoclonal antibody (MAb) is a single type of antibody that is directed against a specific antigenic determinant(epitope). MAbs are immunoglobulins or a fragment of an immunoglobin produced by a single clone of cells. They can be obtained from immortalized B- lymphocytes that are cloned and expanded as continuous cell lines(murine and human monoclonal antibodies) or from rDNA engineered mammalian or bacterial cell lines(engineered monoclonal antibodies). When a foreign body or antigen molecule enters into the body, the immune response begins.
  • 4. 4 PRODUCTION OF MONOCLONALANTIBODY • The establishment of hybridomas and production of MAbs involves the following steps: 1. Immunization 2. Cell fusion 3. Selection of hybridomas 4. Screening the products 5. Cloning and propagation 6. Characterization and storage
  • 6. 6 PRINCIPLE FOR THE CREATION OF HYBRIDOMA CELLS • Hybridoma cells can be produced by the fusion of an antibody producing cell and a myeloma cell. • The mammalian cells can synthesize nucleotides by two pathways:- A)De novo synthesis B)Salvage pathway
  • 7. 77 PRINCIPLES OF HAT SELECTION 1. When the major synthetic pathways are blocked by the folic acid analogue aminopterin, the cell should employ the salvage pathway. This pathway essentially contains the enzyme HGPRT. 2. HGPRT- cells die in the presence of HAT because both the major synthetic pathway and the salvage pathway are blocked virtually. 3. Fusion of the HGPRT- myeloma cells with the HGPRT+ spleen cells allows growth in HAT by incorporating the missing enzyme for the salvage pathway.
  • 9. 99 APPLICATIONS OF MONOCLONALANTIBODIES • Cancer therapy or immunotoxicology • Bone marrow transplantation • Organ transplantation • Protein purification • Disease diagnosis • Gene cloning and expression
  • 10. 1010 LATEST NEWS ON MONOCLONALANTIBODIES • MAb pembrolizumab prolongs the survival in patients with squamous cell carcinoma.(Review on JAN 18,2019) • Tychan has successfully conducted the Phase 1 trial of TY014, a first-in-class monoclonal antibody candidate treatment for yellow fever. (13th DEC 2018) • The meta-analysis of 10 randomized controlled trials has confirmed that human MAb denosumab can more effectively increase the bone mass in patients with low bone mineral density or osteoporosis. Any other biphosphate therapy can not do. (13th DEC 2018) • Sarlumab developed in collaboration with Sanofi and Regeneron Pharmaceuticals, is a MAb targeting interleukin 6 receptors approved by the FDA. (May 2017) • Roche’s Emicizumab, the last approved MAb drug in 2017, targets the coagulation factors IXa and X, thereby promoting the clotting process.
  • 11. 1111 CONCLUSIONS • Pathogens can be detected in a small number of hours sparing few days of culturing of cells. • Since for the human health care, the subject of production and use of antibodies has become a very important area of research, not only for the academic purpose but also for its relevance to industrial growth for diagnosis and therapeutics, MAb can be utilized for it. • MAb technology in conjunction with recombinant DNA technology has successfully led to the reconstruction of chimeric, humanized and fully human antibodies.
  • 12. 1212 REFERENCES • ‘Sambamurthy K., Kar Ashutosh’, “Pharmaceutical Biotechnology” Reprint of First Edition: 2008, New Age International (P) Ltd. Publishers, ISBN:81-224-1911-9, Chapter 1: Immunology and Immunological Preparations, Subunit 5: Antigen Antibodies Reactions and Their Applications (5.1.5 Monoclonal Antibodies), Page 53-58. • Dr. U Satyanarayana, “Biotechnology” Reprint Of First Edition:2010, Books And Allied (P) Ltd., ISBN:81-87134-909, Chapter17 Monoclonal Antibody (Hybridoma Technology) Page- 213-214.