title: Balanced expression of single subunits in a multisubunit protein, achieved by cell fusion of individual transfectants.
abstract: To establish stable cell lines that produce recombinant multisubunit proteins, it is usually necessary to cotransfect cells with several independent gene constructs. Here, we show that a stepwise fusion of individually transfected cells, results in a fused cell-line that secretes a complete multisubunit protein. Functional expression of recombinant multisubunit proteins may require a defined expression ratio between each protein subunit. The cell-fusion technology described allows a predefined expression level of each subunit. Using SIgA as a model protein we demonstrate that the majority of the fused cells inherit the molar expression ratio of the parental transfected cells. These results indicate that simplified screening of clones expressing the expected subunit ratios may be possible using the cell-fusion technology. This technology may therefore be an alternative to generic transfection methods for the establishment of cells that produce multiprotein complexes such as antibodies, receptors, ion channels and other multisubunit proteins.
authors.affiliation: Antibody Design AS, PO Box 190, N-1450 Nesoddtangen, Norway. lars.norderhaug@antibodydesign.com
authors.forename: Lars
authors.initials: L
authors.lastname: Norderhaug
authors.forename: Finn-Eirik
authors.initials: FE
authors.lastname: Johansen
authors.forename: Inger
authors.initials: I
authors.lastname: Sandlie
mesh_terms: D000818:Animals; D016466:CHO Cells; D002459:Cell Fusion / Q000379:methods*; D006224:Cricetinae; D007070:Immunoglobulin A / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D007070:Immunoglobulin A; D021122:Protein Subunits; D011994:Recombinant Proteins