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Monoclonal Antibodies in rDNA technology

To describe about monoclonal Antibodies in hybridoma technology

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Monoclonal Antibodies•
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VIVEKANANDHA ARTS AND SCIENCE COLLEGE FOR WOMEN Veerachipalayam – 637303,Sankagiri, Salem D.t, Tamilnadu, India. Affiliated to Periyar University, Salem; Recognised Under section 2(f) & 12(B) of the UGC Act, 1956. SUBJECT INCHARGE: Dr R. Mythili , Head of the department , Department of Microbiology, Vivekanandha arts and science College for women. SUBMITTED TO: A. Vidhya , Department of Microbiology , Vivekanandha arts and science College for women. DEPARTMENT OF MICROBIOLOGY SUBJECT: Molecular biology and rDNA technology TOPIC: Monoclonal Antibodies in Therapy – vaccines and their applications in animal infection
Monoclonal Antibodies in Therapy VACCINES AND THEIR APPLICATIONS IN ANIMAL INFECTION
Synopsis:  Introduction  What are monoclonal Antibodies  History and development  Production of monoclonal Antibodies  Therapeutic applications  Role of mAbs in vaccine technology  Applications of mABs in animal infection
Introduction: Monoclonal antibodies (mAbs) are one of the most important advances in modern biotechnology and immunotherapy. They are laboratory-produced antibodies that are designed to bind specifically to a single antigen or epitope. Because of their high specificity, monoclonal antibodies are widely used in the treatment of cancers, autoimmune diseases, infectious diseases, and are being increasingly explored in veterinary medicine  Unlike conventional drugs, monoclonal antibodies target only diseased cells or specific molecules, reducing damage to healthy tissues.
Objectives: Understand the concept of monoclonal antibodies. Explain the production of monoclonal antibodies. Describe the mechanism of action of mAbs. Discuss therapeutic applications in animal infections. Explain different types of veterinary vaccines. Compare monoclonal antibodies with vaccines.  Explore future prospects in veterinary immunotherapy.
History of monoclonal Antibodies 1975: Georges Köhler and César Milstein developed Hybridoma Technology. 1984: Nobel Prize awarded. 1986: First therapeutic monoclonal antibody approved.  Today, monoclonal antibodies are widely used in human medicine and are increasingly being developed for veterinary applications.
What are monoclonal Antibodies Monoclonal antibodies are identical antibodies produced from a single clone of B lymphocytes. They recognize one specific antigenic epitope with high specificity. Characteristics Highly specific Uniform structure High affinity Consistent quality
Production of monoclonal Antibodies  Antigen selection | Mouse immunization | Isolation of spleen B cells | Fusion with myeloma cells ( peg) | Hybridoma formation |
Selection of antibodies | Screening ( ELISA ) | Cloning | Purification | Quality control | Therapeutic monoclonal antibiotics
Hybridoma technology  Hybridoma technology is a method used to produce monoclonal antibodies (mAbs)—identical antibodies that recognize a single specific antigen. It was developed by Georges Köhler and César Milstein in 1975.  It was mainly used for production of monoclonal Antibodies in Therapy
Principle of hybridoma technology The technique involves fusing: B-lymphocytes (plasma cells): Produce specific antibodies but have a short lifespan. Myeloma (cancer) cells: Divide indefinitely but do not produce antibodies.  The fused cell, called a hybridoma, combines the ability to produce a specific antibody with unlimited growth.
Steps involved in hybridoma technology Immunization A mouse is immunized with the target antigen. The mouse develops B cells that produce antibodies against the antigen. Isolation of Spleen Cells  Spleen cells containing antibody-producing B lymphocytes are collected.
Fusion Spleen B cells are fused with myeloma cells using polyethylene glycol (PEG). This forms hybridoma cells. Selection in HAT Medium Cells are cultured in HAT (Hypoxanthine-Aminopterin-Thymidine) medium. Unfused myeloma cells die. Unfused B cells die naturally after a few days.  Only hybridoma cells survive and multiply.
Screening Hybridomas producing the desired antibody are identified using tests such as ELISA. Cloning The selected hybridoma is cloned (e.g., by limiting dilution) to obtain a pure cell line.  Production and Purification
Production and Purification Hybridoma cells are cultured in bioreactors (or historically in mouse ascites).  Monoclonal antibodies are harvested and purified.
Hybridoma technology
Types of monoclonal Antibodies  Monoclonal antibodies are classified based on how much of the antibody is derived from mouse or human protein. 1) Murine (-omab) 2) chimeric ( -ximab) 3) Humanized monoclonal antibodies ( -zomab) 4) Fully Humanized monoclonal antibodies (-umab)
Murine Monoclonal Antibodies (-omab)  A murine monoclonal antibody is a lab-made protein created entirely from mouse cells.  Because it is 100% mouse protein, the human body usually rejects it as a foreign substance. Today, it is mostly used in laboratory research and diagnostics rather than as a direct medicine for humans.
Advantages: Easy to produce. High affinity for the target antigen. Disadvantages: Can trigger a human anti-mouse antibody (HAMA) response. Shorter half-life in humans.  Example: Muromonab-CD3 (OKT3).
Chimeric monoclonal Antibodies (-Xiamb)  Chimeric monoclonal antibodies (mAbs) are genetically engineered proteins developed to bridge the gap between early rodent-derived antibodies and modern, fully human therapeutics.  They combine genetic components from two different species—typically a laboratory rodent and a human—to create a functional treatment that is highly compatible with the human body.
Advantages: Less immunogenic than murine antibodies. Longer half-life. Disadvantages: Some immune reactions may still occur. Examples: Rituximab  Infliximab
Humanized Monoclonal Antibodies (-zumab)  Humanized monoclonal antibodies (mAbs) are laboratory-engineered proteins designed to bind to specific targets in the body, such as cancer cells or viruses.  They are constructed by taking antibodies from a non- human species (typically mice or rats) and altering their protein sequences to closely match those found naturally in humans.
Advantages: Very low immunogenicity. High therapeutic effectiveness. Longer duration of action. Disadvantages: More expensive to develop. Examples: Trastuzumab
Fully Human Monoclonal Antibodies (-umab)  Fully human monoclonal antibodies (mAbs) represent the peak of therapeutic antibody engineering.  Unlike humanized antibodies—which still retain tiny fragments of mouse or rat gene sequences in their binding loops—fully human antibodies contain 100% human protein sequences.
Advantages: Lowest risk of immune reactions. Long half-life. Highly effective for long-term therapy. Disadvantages: Complex and costly production. Examples: Adalimumab
Vaccines  Vaccines stimulate the animal’s immune system to produce active immunity and long-term immune memory, whereas monoclonal antibodies provide immediate but temporary protection. Together, they are valuable tools in controlling animal diseases.
 Role of Monoclonal Antibodies in Vaccines  Identify protective antigens for vaccine development.  Test vaccine quality and potency.  Study the immune response after vaccination.  Improve vaccine research and design.  Provide passive protection until vaccine-induced immunity develops.
Types of vaccines in veterinary Live Attenuated Vaccines Examples : Newcastle disease PPR Advantages: Strong immunity Long-lasting protection Disadvantages:
Inactivated Vaccines Examples: Rabies Foot-and-mouth disease Advantages: Safe Stable Disadvantages:  Booster doses required
Subunit Vaccines Contain purified proteins. Advantages: Very safe  Fewer side effects
Recombinant Vaccines Produced using genetic engineering. Examples: Recombinant rabies vaccine Advantages: High safety  Target-specific immunity
mRNA Vaccines Newest vaccine technology. Advantages: Rapid development High precision  Potential future use in veterinary medicine
Applications Cattle CattleBrucellosis Hemorrhagic septicemia Black quarter Poultry Newcastle disease Marek’s disease Infectious bursal disease
Dogs Rabies Canine parvovirus Adenovirus Cats Panleukopenia Calicivirus Swine
Reference  Monoclonal Antibody Applications in Travel Medicine (2024)  Current Review of Monoclonal Antibody Therapeutics in Small Animal Medicine (2025)   Kuby Immunology - Authors: Jenni Punt, Sharon Stranford, Patricia Jones, Judy Owen  Cellular and molecular Immunology – Immunological basis of Monoclonal Antibody
Monoclonal Antibodies in rDNA technology