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Genetic engineering of human lymphocytes for the production of monoclonal antibodies

Genetic engineering of human lymphocytes for the production of monoclonal antibodies The production of human monoclonal antibodies is limited by a lack of good human fusion partners, the short-term stability of most human hybridomas, low fusion frequencies, and low secretion levels. An alternative method of immortalizing human immunoglobulin-secreting lymphocytes is presented here, which avoids many of the drawbacks associated with human hybridoma technology. Briefexposure of cells to a high intensity electric field results in the reversible permeabilization of cell membranes (electroporation). This technique allows the introduction of genes into virtually any cell type in a highly efficient manner. Specific genes or DNA sequences can thus be transferred into cells via the transient pores created in the membrane and, if the sequences are expressed, can yield cells with altered phenotypic properties (i.e., drug resistance). Electroporation of activated human lymphocytes in the presence of DNA sequences coding for immortalization functions and specifically targeted to B cells, would yield, via the stable integration of exogenous DNA into the cellular genome and the subsequent expression of the genes, immortalized, immunoglobulin-secreting lymphocytes, without the needfor a fusion partner. Details of this technique, in addition to its advantages over the commonly used methods for generating hybridomas, are discussed. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Human Antibodies IOS Press

Genetic engineering of human lymphocytes for the production of monoclonal antibodies

Mally, Martin I.
Human Antibodies , Volume 1 (1) – Jan 1, 1990
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References (51)

Publisher
IOS Press
Copyright
Copyright © 1990 by IOS Press, Inc
ISSN
1093-2607
eISSN
1875-869X
DOI
10.3233/HAB-1990-1106
Publisher site
See Article on Publisher Site

Abstract

The production of human monoclonal antibodies is limited by a lack of good human fusion partners, the short-term stability of most human hybridomas, low fusion frequencies, and low secretion levels. An alternative method of immortalizing human immunoglobulin-secreting lymphocytes is presented here, which avoids many of the drawbacks associated with human hybridoma technology. Briefexposure of cells to a high intensity electric field results in the reversible permeabilization of cell membranes (electroporation). This technique allows the introduction of genes into virtually any cell type in a highly efficient manner. Specific genes or DNA sequences can thus be transferred into cells via the transient pores created in the membrane and, if the sequences are expressed, can yield cells with altered phenotypic properties (i.e., drug resistance). Electroporation of activated human lymphocytes in the presence of DNA sequences coding for immortalization functions and specifically targeted to B cells, would yield, via the stable integration of exogenous DNA into the cellular genome and the subsequent expression of the genes, immortalized, immunoglobulin-secreting lymphocytes, without the needfor a fusion partner. Details of this technique, in addition to its advantages over the commonly used methods for generating hybridomas, are discussed.

Journal

Human Antibodies – IOS Press

Published: Jan 1, 1990

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