Monoclonal Antibodies

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Presentation transcript:

Monoclonal Antibodies

CELLS OF THE IMMUNE SYSTEM

IMMUNOTHERAPY Treatment of the disease by Inducing, Enhancing or Suppressing the Immune System. Active Immunotherapy: - It stimulates the body’s own immune system to fight the disease. Passive Immunotherapy: - It does not rely on the body to attack the disease, instead they use the immune system components ( such as antibodies) created outside the body.

ANTIBODIES STRUCTURE CLASS

PRODUCTION OF MONOCLONAL ANTIBODY HYBRIDOMA TECHNOLOGY

Ab titre reached in Serum PRODUCTION OF MONOCLONAL ANTIBODY HYBRIDOMA TECHNOLOGY Step 1: - Immunization Of Mice & Selection Of Mouse Donor For Generation Of Hybridoma cells ANTIGEN ( Intact cell/ Whole cell membrane/ micro-organisms ) + ADJUVANT (emulsification) Ab titre reached in Serum Spleen removed (source of cells)

PRODUCTION OF MONOCLONAL ANTIBODY HYBRIDOMA TECHNOLOGY Step 2: - Screening Of Mice For Antibody Production After several weeks of immunization Serum Antibody Titre Determined (Technique: - ELISA / Flow cytometery) Titre High Titre too low 2 weeks BOOST (Pure antigen) BOOST (Pure antigen)

+ PRODUCTION OF MONOCLONAL ANTIBODY 8 - Azaguanine Myeloma Cells HYBRIDOMA TECHNOLOGY Step 3: - Preparation of Myeloma Cells + 8 - Azaguanine Myeloma Cells Immortal Tumor Of Lymphocytes Myeloma Cells HGPRT- High Viability & Rapid Growth

PRODUCTION OF MONOCLONAL ANTIBODY HYBRIDOMA TECHNOLOGY Step 4: - Fusion of Myeloma Cells with Immune Spleen Cells & Selection of Hybridoma Cells PEG FUSION MYELOMA CELLS SPLEEN CELLS Feeder Cells Growth Medium Plating of Cells in HAT selective Medium Scanning of Viable Hybridomas HYBRIDOMA CELLS ELISA PLATE HAT Medium

PRODUCTION OF MONOCLONAL ANTIBODY HYBRIDOMA TECHNOLOGY Step 4: - Cloning of Hybridoma Cell Lines by “ Limiting Dilution” or Expansion A. Clone Each +ve Culture B. Test Each Supernatant for Antibodies C. Expand +ve Clones Tissue Culture Method Mouse Ascites Method

PRODUCTION OF MONOCLONAL ANTIBODY HYBRIDOMA TECHNOLOGY