Monoclonal antibodies preparatation and applicatation

monoclonal antibodies preparatation and applicatations

1. MONOCLONAL ANTIBODIES PREPARATION & APPLICATIONS Prepared By, Sujitha Mary MPHARM ST.Joseph’s College Of Pharmacy 1

2. An antibody is a protein used by the immune system to identify and neutralize foreign objects like bacteria and viruses.Each antibody recognizes a specific antigen unique to its target. Monoclonal antibodies ( mAb ) are antibodies that are identical because they were produced by one type of immune cell;all clones of a single parent cell. Basic Structure of antibody 2

3. Characters of Monoclonal Antibodies Monoclonal Antibodies are single type of antibody that are identical and are directed against a specific antigen. These antibodies are produced by B-cell clones of a single parent or a single hybridoma cell line. A hybridoma cell line is formed by the fusion of one B –cell lymphocyte with a myeloma cell. Some myeloma cells synthesize single mAB antibodies naturally. 3

4. Advantages of using mAB mAB are cheaper to develop than conventional drugs because it is based on tested technology. Sideeffects can be treated and reduced by using mice-human hybrid cells or by using fraction of antibodies. They bind to specific diseased or damaged cells needing treatment. They treat a wide range of conditions . 4

5. Disadvantages Time consuming project-between 6-9months. Hybridoma culture may be subject to contamination. System is only well developed for limited animal and not for other animals. Morethan 99% of the cells do not survive during the fusion process . Possibility of generating immunogenicity. 5

6. Production of Monoclonal antibodies A ) Hybridoma Technology :- Hybridoma technique for monoclonal antibody production was discovered by, George.J.K and Cesar Milstein in 1975 Steps 1. Immunisation of mice with a specific antigen 2. Booster dose of antigen 3. Measurement of antibody synthesis 4. Collection of splenocytes 5. Myeloma cells selection 6. Fusion of cells for production of hybridoma cells 7. Selection of Hybridoma cells 8. Selection of specific hybridoma cells 9. Checking for monoclonality 10. Selection of specific hybridoma 11. Propogation of selected hbridoma 12. Purification of Antibodies monoclonal 6

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14. B) By rDNA technology 14

15. Applications of Monoclonal Antibodies DIAGNOSTIC APPLICATION THERAPEUTIC APPLICATION PROTEIN PURIFICATION MISCELLANEOUS APPLICATIONS 15

16. DIAGNOSTIC APPLICATION :- a) mAb’s in biochemical analysis :- Used in ELISA & RIA These assays measure circulating concentration of hormones( insulin,growth hormones) & cell products (blood group antigens,blood clotting factor) b) Pregnancy kits :- The hCG card pregnancy test is a rapid chromatographic immunoassay for the quantitative detection of human chorionic gonadotropin. c)For detecting hormonal disorders d) Infectious diseases :- Detection of antigen of Nisseria gonorrhoea & Herpes simplex virus. e)Radiolabelled mAbs are used in the diagnostic imaging. f) Cardiovascular disease :- mAb labelled with radioisotopic Indium chloride is used for detecting site of MI Febrin specific mAb can be used for detection of clots in thigh,pelvis &knee regions. g)It has been possible to detect certain types of cancers ( lung,ovarian ) at early stages by employing mAbs 16

17. THERAPEUTIC APPLICTIONS :- a)Used for enhancing immune formation of host. b)In the treatment of cancer:- Ab will binds with the cancer cells and destroy them by Ab -dependent cell mediated cytotoxicity & phagocytosis of cancer cells by reticulo endothelial system. c)In the organ transplantation:- OKT3 specifically directed against CD3 antigen of T-lymphocyte is used in renal & bone marrow transplantation. d)As immunotoxins :- Toxins coupled with mAbs to form immunotoxin ad used in therapy eg:Diphtheria toxins,pseudomonas endotoxins. e)In drug delivery:- mAbs can be conjugated to cytotoxic drugs & can deliver drugs to cancer cells while minimizing damage to normal cells. 17

18. PROTEIN PURIFICATION :- mAb can be produced for any protein and the so produced mAb can be used for purification of protein against it was raised. mAb columns can be prepared by, coupling them to cyanogen bromide activated sepharose . The immobilised mAbs in this manner are very useful for purification of proteins by immunoaffinity method. 18

19. MISCELLANEOUS APPLICATION :- a)Catalytic mAb ( Abzymes ):- The Ab enzymes regarded as abzymes,are catalytic Ab. There is difference in the Ab recognition of an antigen & enzyme recognition of substances. While the Ab recognize in ground state, enzymes recognize in transition state. b)Autoantibody fingerprinting:- Auto antibodies are produced after birth & reach maximum in no. 2yrs,and they remain constant for later part of life. Used for the detection & identification of indivituals . 19

20. Reference:- Shivanand.P (2010). Hybridoma technology for production of monoclonal antibodies,International journel of pharmaceutical sciences review and research vol.1. Lachman / Liberman’s the theory & practice of industrial pharmacy,Editors :; Roop khar,SP Vyas,Farhan Ahmad,Gaurav Jain 20