Monoclonal by akashyap

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									.Monoclonal Antibodies
Cell Culture Myeloma line

Fuse in Polyethylene Glycol

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Myeloma Cells (HPRT -)

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Speen Cells (HPRT-)

HAT Medium
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Select Hybrid Cells

Assay for Antibody
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Clone

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Freeze

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Assay for Antibody
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Reclone

Freeze

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Analyze t 0 select

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vanants

Grow"

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Thaw
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Mif
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Antibody

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Antibody

Fig. 1. Production monoclonal ntibodies. of a

Monoclonal Antibodies
. Discovered by G.Kohler and Milstein in 1975.Technique is hybridoma
technology. Conventional antisera is heterogeneous mixture of antibodies of different specificities, affinities and classes. Monoclonal antibodies produced by single clone of B cells directed against single antigenic determinant. Uniform, highly specific and produced in large quantities.

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How to produce them
I. Mouse is immunized with specific antigen to produce antibodies. 2. Mouse spleen is removed; suspension is made which contains B cells that can produce antibodies to injected antigen. Spleen cells have enzyme HPRT (hypoxanthine phosphorybosyl transferase), but myaloma cells lack this enzyme. 3. Spleen cells mixed with cultured myeloma cells to form hybrids, hybrid can grow continuously & produce large amount of antibodies.( property of spleen cell to produce antibodies and myaloma cells to multiply continuously) 4. Fusion occurs in presence of polyethylene glycol. s. Hybrids made to grow in HAT ( hypoxanthine, aminopterin and thymidine) medium, proliferate to hybridomas ,desired hybrids are made to grow which will produce monoclonal antibodies. 6. Unfused spleen cells can not multiply for lond and unfused myaloma cells killed by aminopterin. 7. Hybrids can be grown as turnours in peritoneal cavity of mice or frozen.

Disadvantage of mouse monoclonal antibodies
Unsuitable for human therapeutic use, induce strong antimouse immune response. Human monoclonal anti bodies produced by introducing genes for particular Antibodies fragment with bacteriophage genes, bacteriophage infects bacteria and multiplies ,produces large amount of genes, which can be harvested.

Uses
1. Diagnostic use- For identification of bacterial ,viral and other antigens, by tagging then with enzymes or florescent dye. Eg Direct florescence or ELISA. 2. Therapeutic use - Cancer ,immunodeficiency, viral infections. 3. Clinical imaging- tag them with radioactive substance to spot turnours


								
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