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Similar to Monoclonal antibody, Application of Monoclonal Antibody, Uses
Monoclonal antibody, Application of Monoclonal Antibody, Uses
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- 2. 2 Antigen • Anysubstance that induces the immune system to produce antibodies against it is called an antigen. Any foreign materials , such as pathogens (bacteria and viruses), chemicals, toxins, and pollens • An antigen (Ag) is a molecule or molecular substance such as may be present on the outside of a body, that can be bound by an antigen-specific antibody or B-cell antigen receptor. The presence of antigens in the body normally triggers an immune response. • An epitope refers to the specific target against which an individual antibody binds. • When an antibody binds to a protein, it isn’t binding to the entire full-length protein. Instead, it is binding to a to a segment of that protein known as an epitope. In general, an epitope is approximately five or six amino acids in length.
- 3. 3 B cellsare formed from multipotent hematopoietic stem cells (HSCs) in the bone marrow. B cells are a type of lymphocyte that are responsible for the humoral immunity component of the adaptive immune system. These white blood cells produce antibodies, which play a key part in immunity. Each B cell contains a single round nucleus B Cells
- 4. 4 Antibody : •It is specialized Glycoprotein produced from activated B cell (Plasma cell) in response to an antigen that trigger its production. • An antibody is a protein component of the immune system that circulates in the blood, recognizes foreign substances like bacteria and viruses, and neutralizes them. After exposure to a foreign substance, called an antigen, antibodies continue to circulate in the blood, providing protection against future exposures to that antigen. • An antibody (Ab), also known as an immunoglobulin (Ig), it is a large, Y-shaped protein used by the immune system to identify and neutralize foreign objects such as pathogenic bacteria and viruses.
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- 6. 6 Monoclonal Antibody i.A class of highly specific antibodies produced by the single clone of hybrid cell ii. Produced by fusion a B cell secreting the desired antibody with a tumor cell ( myeloma cell ) Capable of growing indefinitely iii. Fused cell called Hybridoma iv. Monoclonal antibodies all have identical antigen-binding sites Bind to the same epitope with same affinity Same antibody class
- 7. 7 Polyclonal Monoclonal Produced byMany B cell clones A single B cell clone Bind to: Multiple epitopes of all antigens used in the immunization A single epitope of a single antigen Antibody class A mixture of different Ab classes (isotopes) All of single Ab class Ag- binding sites: Different antigen-binding site All Abs have the same antigen binding site Cost Less expensive More expensive
- 8. 8 • Types ofmonoclonal antibodies Naked Monoclonal Antibody : Those without any drug or radioactive material attached to them. Marks the cells from immune system Attach to receptor - block binding of growth factor E.g : 1) Trastuzumab - for advanced breast cancer 2) Rituximab : For B cell non Hodgkin lymphoma 3) Cetuximab : for metastatic colorectal cancer . Conjugated / Loaded monoclonal antibody Coupled with drug / toxins/ radioactive atoms
- 9. 9 Chemo-labeled antibodies: MAbsConjugated with chemo therapeutic agents Eg: Brentuximab vedotin Brentuximab vedotin attached to a chemo drug target the CD30 antigen ( present on T and B cells) in treatment of Hodgkin lymphoma Immune-Toxins: Conjugated with toxins Eg : Denileukin diftitox Used to treat some cancer (cutaneous T-cell lymphoma and many other) Consist of IL-2 protein attaches to toxin (derived from the germ causing diphtheria) IL-2 normally attached to cell that express the CD25 antigen and thus helps in delivering the toxins to these cell
- 10. 10 Production ofMonoclonal Antibodies. Production in animals Mouse ascites method • Hybridoma cells injected in mouse • Produce ascites (Excessive Abdominal fluid) • Fluid contain high concentration of Abs • No further concentration required • Purification required • Easy and inexpensive
- 11. 11 Production incall cultured ( In - vitro) Batch tissue culture method • Grow Hybridoma cells in batch • Purify MAbs from the culture media • Fetal bovine serum commonly use • Low concentration • Semi permeable membrane based system
- 12. 12 Production of MonoclonalAntibodies. Step 1 – Immunization of Mouse
- 13. 13 Step 2 –Screening of Mice For Antibody Production
- 14. 14 Step 3 –Isolation of antibody producing spleen cell
- 15. 15 Step 4 –Production of Hybridoma Cell
- 16. 16 Step 5 –Screening of Hybridomas
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- 18. 18 Step 6 –Culturing Hybridoma cell – Monoclones Production
- 19. 19 Step 7- Screeningfor Desired antibodies
- 20. 20 Step 8 –Selection and culture of screened antibody
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- 22. 22 Route ofadministration Subcutaneously : ( Rituximab, Trastuzumab, adlimumab) Intramuscularly : ( Palvizumab) Intravenously : Preferred because of 100% Bioavailability Route of elimination of antibodies Via uptake and catabolism Half-life - Chimeric : 4-15 days - Humanized 3-24 days - Recombinant human : 11-24 days
- 23. 23 How do monoclonalantibody drugs work? Flagging cancer cells. Triggering cell-membrane destruction Blocking cell growth. Preventing blood vessel growth Blocking immune system inhibitors. Directly attacking cancer cells Delivering chemotherapy
- 24. 24 USES OF MABSSIDE EFFECTS OF MABS I. Cancer II. Rheumatoid arthritis III. Multiple sclerosis IV. Cardiovascular disease V. Systemic lupus erythematosus VI. Crohn's disease VII. Ulcerative colitis VIII. Psoriasis IX. Transplant rejection I. Allergic reactions II. Chills III. Weakness IV. Diarrhea V. Nausea VI. Vomiting VII. Rash VIII. Itching IX. High blood glucose levels
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- 27. 27 Application of MonoclonalAntibody Diagnostic tools in research and laboratory Different technique in which MAbs are used include Western blot, ELISA, radioimmune Assay ( RIA), Fluorescence microscopy Diagnostic application MAbs is used to detect pregnancy as early as a week or two after conception by reacting with human chorionic gonadotropin HIV diagnostic kits Rapid diagnosis of hepatitis, influenza, herpes Identification and characterization of tumor specific antigen
- 28. 28 Fight againstbio terrorism Inhalation anthrax ( Potent biological terrorism) is caused by breathing the bacterial spores of bacillus anthracites. Raxibacumab injection is used to treat infectious inhalational anthrax when alternative therapies have failed.
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