Antibodies, A Laboratory Manual, Second Edition, Edited by Edward A. Greenfield



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Preparing Live Cells for Immunization

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Generating monoclonal antibodies against cell-surface proteins (membrane proteins) is very challenging because membrane and membrane-associated proteins often lose their native conformation during the purification process. This also makes fusion screening very difficult. One of the widely used techniques to overcome this issue is to overexpress the target protein in HEK 293T cells and then immunize the host with these cells (Dreyer et al. 2010). This facilitates presentation of the target protein in its native conformation to the host immune system. Screening hybridomas using the same cells and counterscreening them on untransfected cells will enable the selection of specific hybridomas.

Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition

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