title: High-yield production of recombinant antibody fragments in HEK-293 cells using sodium butyrate.
abstract: To develop new recombinant monoclonal antibody fragments for therapy and imaging, it is indispensable to have a simple and easy procedure to handle the eukaryotic expression system for production of proteins in high amounts. Gene amplification techniques such as the dehydrofolate reductase (DHFR) system in Chinese hamster ovary cells or the glutamine synthase system in myeloma cells have a couple of disadvantages. The selection procedure is complex, time-consuming, and not fruitful in all cases. The toxic drug methotrexate (for the DHFR system) can increase the production rate but decreases the specific growth rate of the cells. The production rate is not always stable over a long-term cultivation period. To overcome these problems, we are using stably transfected human embryonic kidney (HEK-293) cells in combination with an efficient screening method. Sodium butyrate can increase the expression of recombinant antibody fragments in the transfectomas up to 500 micrograms/4.2 x 10(7) cells/24 h corresponding to 175 micrograms/mL culture medium. This strategy allows a rapid development of new recombinant monoclonal antibody fragments and allows one to proceed rapidly to in vivo testing.
authors.affiliation: ETH-PSI-USZ Villigen-PSI, Switzerland. juergen.gruenberg@psi.ch
authors.forename: Jürgen
authors.initials: J
authors.lastname: Grünberg
authors.forename: Karin
authors.initials: K
authors.lastname: Knogler
authors.forename: Robert
authors.initials: R
authors.lastname: Waibel
authors.forename: Ilse
authors.initials: I
authors.lastname: Novak-Hofer
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000302:isolation & purification; D002087:Butyrates / Q000494:pharmacology*; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D005786:Gene Expression Regulation / Q000187:drug effects*; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D007668:Kidney / Q000187:drug effects* / Q000196:embryology / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014162:Transfection / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D002087:Butyrates; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins