Monoclonal antibodies

complete description about monoclonal antibodies...

1. Monoclonal Antibodies GROUP: 09 PRESENTED TO: MA’AM HUMA DILSHAD

2. INTRODUCTION & DISCOVERY IMMUNE SYSTEM!!! Latin term "IMMUNIS" means EXEMPT, referring to protection against foreign agents. An integrated body system of organs, tissues, cells & cell products that differentiates self from non-self & neutralizes potentially pathogenic organisms.

3. CELLS OF IMMUNE SYSTEM

4. WHAT ARE ANTIBODIES OR IMMUNOGLOBULIN ?

5. ANTIBODIES...

6. DIFFERENCE!!!

7. DISCOVERY!!! The idea of a "magic bullet" was first proposed by Paul Ehrlich at beginning of 20th century. He postulated that if a compound could be made that selectively targeted a disease-causing organism, then a toxin for that organism could be delivered along with agent of selectivity.

8. DISCOVERY!!! George Kohler & Cesar Milstein in 1975 shared the Nobel Prize in Physiology or Medicine in 1984 for discovery of hybridoma technology.

9. The structure of antibodies

10. Definitions

11. THE TYPES OF MONO-CLONAL ANTIBODIES 1- According To Evolution 2- According to design

12. ‘Naked’ Monoclonal Antibodies

13. Conjugated/ labelled/ loaded Monoclonal A ntibodies Conjugated monoclonal antibodies are those joined to a chemotherapy drug, radioactive particle, or cancer cell killing agent. Gemtuzumab ozogamicin ( Mylotarg )

14. Immune-Toxin Monoclonal Antibody

15. Principles in Hybridoma technology

16. Cont’d

17. Process by which bulk quantities of targeted antibodies against a specific antigen are produced.Monoclonal antibodies are produced via multiple/identical copies of a certain cell called a hybridoma .

18. HGPRT= H ypoxanthineguanine phosphoribosyltransferase

19. PRODUCTION OF MONOCLONAL ANTIBODY INVOLVE’S:

20. Step 1: - Immunization Of Mice & Selection Of Mouse Donor For Generation Of Hybridoma cells ANTIGEN ( Intact cell/ Whole cell membrane/ micro-organisms ) + ADJUVANT (emulsification) The first step in making a hybridoma is to generate antibody producing B cells.This is done by immunizing a mouse against the antigen of interest.Intraperitoneal (IP)injections are the most common method for delivering the antigens in to mice.

21. Step 2: - Screening Of Mice For Antibody Production After several weeks of immunization Screening Of Mice For Antibody Production It must be determined if the mouse is producing antibodies of interest.Test bleeds are performed and examined for the presence of antibodies. If the host is producing the desired antibody,the spleen is removed and dissociated in culture medium to release the resident B cells.The culture medium also includes cells from a special mouse myeloma cell line.These tumor cells can divide indefinitely,but do not produce antibodies

22. Immortal Tumor Of Lymphocytes + 8 - Azaguanine Myeloma Cells High Viability & Rapid Growth HGPRT Myeloma Cells Step 3: - Preparation of Myeloma Cells

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

24. The next step is to separate the fused hybridoma cells from the unfused B cells and myeloma cells.Unfused B cells will die because they lack the ability to survive in culture. STEP 5:- SEPARATION OF FUSED HYBRIDOMA CELL AND SCREENING Surviving hybridomas are separated and individually cultured—one cell per well.These cells are described as clonal cultures because all of the cells from each well are derived from a single cell,and are therefore identical,or clonal.

25. After culturing for a few weeks,when the growing cells can be seen,the culture fluid can be screened for the presence of desirable antibodies.Screening is the most labor-intensive step in the production of monoclonal antibodies since fusion can result in thousands of individual hybridoma colonies.Note that different antibodies may react with different epitopes on the same antigen. SCREENING

26. Step 6: - 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 Finally,clones that produce the desired antibody are grown in mass culture and/or frozen for storage.

27. DIAGNOSTIC APPLICATION OF MONOCLONAL ANTIBODIES

28. Diagnosis of HIV Infection

29. Pregnancy Tests

30. Pregnancy Tests

31. Purification of proteins

32. Therapeutic Uses

33. Therapeutic Uses

34. Therapeutic Uses

35. Therapeutic Uses

36. New Research

37. The ESK1 monoclonal antibody was engineered to recognize WT1 peptides brought to the surface of cancer cells.

38. SIDE EFFECTS OF MONOCLONAL ANTIBODIES:

39. CONT;

40. SERIOUS SIDE EFFECTS

41. Advantages

42. Dis- Advantages

43. Research 1

44. Research 2