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    Production of Monoclonal Antibodies

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    Mudit Misra
    Monoclonal antibodies are homogeneous populations of antibody molecules derived from a single antibody producing cell that are identical and of the same specificity for a given epitope. Mice are immunized with antigens greater than 3,000-5,000 Da or linked to carriers. Lymphocytes are then fused with myeloma cells using polyethylene glycol to promote membrane fusion. The cells are cultured in HAT medium which lacks the enzyme HGPRT, causing non-fused cells to die while hybridoma cells survive and can be screened for antibody production by ELISA, RIA, or solid phase immunoassay.

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    Production of Monoclonal Antibodies

    Uploaded by

    Mudit Misra
    25 views8 pages
    Monoclonal antibodies are homogeneous populations of antibody molecules derived from a single antibody producing cell that are identical and of the same specificity for a given epitope. Mice are immunized with antigens greater than 3,000-5,000 Da or linked to carriers. Lymphocytes are then fused with myeloma cells using polyethylene glycol to promote membrane fusion. The cells are cultured in HAT medium which lacks the enzyme HGPRT, causing non-fused cells to die while hybridoma cells survive and can be screened for antibody production by ELISA, RIA, or solid phase immunoassay.

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    Monoclonal antibodies are homogeneous populations of antibody molecules derived from a single antibody producing cell that are identical and of the same specificity for a given epitope. Mice are immunized with antigens greater than 3,000-5,000 Da or linked to carriers. Lymphocytes are then fused with myeloma cells using polyethylene glycol to promote membrane fusion. The cells are cultured in HAT medium which lacks the enzyme HGPRT, causing non-fused cells to die while hybridoma cells survive and can be screened for antibody production by ELISA, RIA, or solid phase immunoassay.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    25 views8 pages

    Production of Monoclonal Antibodies

    Uploaded by

    Mudit Misra
    Monoclonal antibodies are homogeneous populations of antibody molecules derived from a single antibody producing cell that are identical and of the same specificity for a given epitope. Mice are immunized with antigens greater than 3,000-5,000 Da or linked to carriers. Lymphocytes are then fused with myeloma cells using polyethylene glycol to promote membrane fusion. The cells are cultured in HAT medium which lacks the enzyme HGPRT, causing non-fused cells to die while hybridoma cells survive and can be screened for antibody production by ELISA, RIA, or solid phase immunoassay.

    Copyright:

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    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 8

    ABHILASHA TIWARI ANKITA DWIVEDI JASMINE KAUR ARIJIT BISWAS

    y Monoclonal anitbodies are homogeneous

    population of antibody molecules,derived from a single antibody producing cell,in which all antibodies are identical and of the same precise specificity for a given epitope.

    Immunisation

    Mice are immunised with Ag


    Antigen must be greater than 3000-5000 Da or linked to a hapten or a carrier protein

    Spleen cell,lymphocytes suspension is obtained .

    Fusion

    Lymphocytes are fused with myeloma cells

    Myeloma cells are malignant lymphocytes . Cells are fused using PEG(polyethyleneglycol) to promote membrane fusion ..

    Culture in HAT
    T e e i i ic t e c lt re is

    .
    ec tai s . yp xa t i e,a i pteri ,t y i i e .

    Myel

    a cells are e gi eere t be eficie t i a e zy e yp xa t i e g a i e p tra sferase(HGPRT) ..

    sp

    rib syl

    He ce t e

    yel

    a cells

    t s rvive i c lt re

    less t is e zy e is a

    e t t e

    e ia

    O ly ybri cells s rvive

    Splee are s

    rt live i.e, t eir life spa is 7-10 ays,s t ey ie gra

    HAT kills

    yel

    a cells ..

    ally ..

    ..

    The cells are cultured in HAT medium which is lacking the enzyme HGPRT

    The powerful toxins of the media block the metabolic pathway Denovo pathway..essential for purine metabolismin DNA synthesis

    Fused cells survive

    Hybridoma cells consisting of immortalized plasma cells fused with B cells will survive in the absence of supplemented HGPRT in culture medium

    Non fused, - HGPRT plasma cells and all non fused B cells die soon.

    Screening
    y ELISA y RIA

    .
    ..

    y SOLID PHASE IMMUNOASSAY

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