monoclonal antibodies

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Monoclonal antibodies : 

Monoclonal antibodies PRESENTED BY:- SANKET B. REKHAWAR DEPARTMENT OF PHARMACEUTICS, S.V.E.TRUST’S COLLEGE OF PHARMACY, HUMNABAD.

Monoclonal antibodies : 

Monoclonal antibodies

Introduction : 

Introduction Cancer chemotherapy drugs are neither specific, i.e., they do not act exclusively on the metabolic pathways of cancer cells, nor are they targeted solely toward cancer cells. However, recent research has begun to address that Improved delivery of chemotherapeutic agents to tumor tissue in man appears to be an achievable goal in the next decade. Monoclonal antibodies directed against tumor-associated antigens have the potential to achieve major advances in targeted drug delivery.

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Monoclonal antibodies may have direct antitumor effects, or they can be used as “homing devices” when attached to a payload and can guide diagnostic or therapeutic agents to the targeted tissues. Monoclonal antibodies may also be bound to the surfaces of macromolecular carriers, such as liposomes. The liposomes can then be loaded with toxin, or a chemotherapeutic agent. The possible advantages of this are substantial. Combinations of McAb can be coupled with the same liposome, and may result in a much greater binding affinity to the targeted tumor.

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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. Polyclonal antibodies are antibodies that are derived from different cell lines. Isotypes According to differences in their heavy chain constant domains, immunoglobulins are grouped into five classes, or isotypes: IgG, IgA, IgM, IgD, and IgE. IgG: IgG1 (66%), IgG2 (23%), IgG3 (7%) and IgG4 (4%) , blood and tissue liquid. IgA:IgA1 (90%) and IgA2 (10%), stomach and intestines

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IgM: normally pentamer, occasionally hexamer, multiple immunoglobins linked with disulfide bonds IgD:1% of proteins in the plasma membranes of B-lymphocytes, function unknown . IgE: on the surface of plasma membrane of mast cells, play a role in immediate hypersensitive and defensive for parasite.

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The structure of antibodies http://www.path.cam.ac.uk/~mrc7/igs/mikeimages.html

History of Monoclonal antibodies development : 

History of Monoclonal antibodies development 1890 Von Behring and Kitasato discovered the serum of vaccinated persons contained certain substances, termed antibodies 1900 Ehrlich proposed the “ side-chain theory” 1955 Jerne postulated natural selection theory. Frank Macfarlane Burnet expended. Almost the same time, Porter isolated fragment of antigen binding (Fab) and fragment crystalline (Fc) from rabbit y-globulin. 1964 Littlefield developed a way to isolate hybrid cells from 2 parent cell lines using the hypoxanthine-aminopterin-thymidine (HAT) selection media. 1975 Kohler and Milstein provided the most outstanding proof of the clonal selection theory by fusion of normal and malignant cells. 1990 Milstein produced the first monoclonal antibodies.

MERITS : 

MERITS Highly applicable to treat carcinomas Higher specificity Selective delivery to target site Protect enzymatic degradation Large number of drugs can be used for targeting. Damage to normal cell can be prevented. DEMERITS Determination of nature & types of antigen on target cell. Lack of in vivo selectivity The mechanism of anti-proliferation on cells cycle, rather than specific toxicity directed towards particular cancer cell Host toxicity: treatment discontinued, most of them had bad side-effects, such as no appetites, omit, loss of hair.

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Monoclonal antibodies for cancer treatment Three mechanisms that could be responsible for the cancer treatment. mAbs act directly when binding to a cancer specific antigens and induce immunological response to cancer cells. Such as inducing cancer cell apoptosis, inhibiting growth, or interfering with a key function. mAbs was modified for delivery of a toxin, radioisotope, cytokine or other active conjugates. it is also possible to design bispecific antibodies that can bind both to target antigen and to a conjugate or effector cell

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mAbs treatment for cancer cells ADEPT, antibody directed enzyme prodrug therapy; ADCC, antibody dependent cell-mediated cytotoxicity; CDC, complement dependent cytotoxicity; MAb, monoclonal antibody; scFv, single-chain Fv fragment.

METHOD OF PREPERATION : 

METHOD OF PREPERATION Mouse is immunized with antigen of interest. B-lymphocytes from the immunized animals spleen or lymph nodes are harvested into single cell suspension. These cells are fused with myeloma (tumor cells) of the same species. Myeloma cells are immortal and have cytoplasmic machinary to survive longer. They contain enzyme called as hypoxanthine guanine phosporibosyl transferase (HGPRT). Fusion is usually accomplished in PEG medium, which triggers fusion. The suspension of cells is disturbed into the bores of micro liter plates in a medium (tissue culture medium) containing hypoxanthine aminopterine thymidine (HAT) medium where only fused cells (hybridoma) survive due the presence of HGPRT.

Applications of Monoclonal Antibodies : 

Applications of Monoclonal Antibodies To study cell – cell interaction. To prepare viral vaccine (by identifying viral antigen). To study ABO and rare blood group. To detect tumor associated antigen. To deliver cytotoxic drugs. In Heart and liver transplant rejection. To locate the tumor within body. Used in enzyme purification .

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Also used in radioimmunoassay. Diagnostic Applications: Biosensors & Microarrays Therapeutic Applications: Transplant rejection Muronomab-CD3Cardiovascular disease - Abciximab Cancer - RituximabInflammatory disease - Infliximab Clinical Applications: Purification of drugs, Imaging the target.

References : 

References Vyas SP, Khar RK. Targeted And Controlled Drug Delivery: Novel Carrier Systems, New Delhi, CBS publisher, 2004, 387-413. Jain NK, Controlled And Novel Drug Delivery, New Delhi, CBS publishers, 2004, 256-281. Gilbert s Banker. Modern Pharmaceutics. 4 th edition Resealed Erythrocytes: An Effective Tool in Drug Targeting Jagadale V.L., Aloorkar N.H., Dohe G.H. and Gehlot M.V. Indian J.Pharm. Educ. Res. 43(4), Oct, -Dec, 2009.p 375-383.

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Thank you

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