Monoclonal antibodies

Monoclonal antibodies

1. MONOCLONAL ANTIBODIES

2. Monoclonal antibodies are monospecific , made by Identical immune cells that are all clones of unique parent cell. Have monovalent affinity, They bind to same epitope. MONOCLONAL ANTIBODIES

3. Immunization of mouse Fusion of myeloma cells and immune spleen cells Culturing in HAT media Separation of hybridoma cells Cloning of hybridoma cells PRODUCTION OF MONOCLONAL ANTIBODIES

4. Hybidoma Technology

5. Cell ,cell debris ,lipids, clotted materials are first removed by filtration. Charged impurities such nucleic acids and endotoxins seperated by ion exchange chromatography. PURIFICATION TECHNIQUES

6. PURIFICATION TECHNIQUES Quick or single step method is Protein A/G affinity chromatograhy.it is problematic for antibodies which can easily damaged. Gel electrophoresis Capillary electrophoresis

7. MURINE ANTIBODIES : Whole antibody is of murine origin. CHIMERIC ANTIBODIES : Murine variable region fused onto human C region. HUMANIZED ANTIBODIES : By grafting murine hyper variable amino acid domains into human antibody. TYPES OF MONOCLONAL ANTIBODIES

8. Diagnostic Applications Biosensors & Microarrays Therapeutic Applications Cancer , Transplant rejection, Infectious diseases Clinical Applications Purification of drugs , Imaging the target Future Applications Fight against Bioterrorism APPLICATIONS

9. NAME TYPE TARGET APPLICATION OKT3 Murine CD3 molecules Acute rejection of organ INFLIXIMAB Chimeric Inhibits TNF-a Rheumatoid arthritis OMALIZUMAB Humanized IgE Moderate to serve allergic asthma PALIVIZUMAB Humanized Inhibits an RSV fusion protein RSV infection in children EXAMPLES OF MONOCLONAL ANTIBODIES

10. When monoclonal antibody attaches to cancer cell : Make the cancer cell more visible to the immune system Rituximab drug attaches to specific protein CD20 found on B cells makes cell more visible to immune system Block growth signals Cetuximab attaches to growth factor receptors , blocking its signal and suppress cancer growth MONOCLONAL ANTIBODIES IN CANCER TREATMENT

11. Stop new blood vessel from forming Bevacizumab blocks growth signal which attract blood vessels then tumor shrinks. Deliver radiation to cancer cells By combining radioactive particle with monoclonal antibodies, radiation are deliver to cancer cells only , cancer cell dies. CANCER TREATMENT