EASE vectors for rapid stable expression of recombinant antibodies.
Over the past 10 years, monoclonal antibodies and antibody fragments have become an increasingly important source of therapeutic molecules in the biotechnology industry. Drug development strategies rely on screening large numbers of candidate molecules in search of an optimized drug candidate. This strategy requires efficient production of ten to a few hundred milligrams of candidate molecules for screening in bioassays and animal models. Typically, this amount of recombinant protein expression involves large numbers of transient transfections or cloning of a recombinant cell line. Both of these approaches are time-consuming and labor-intensive. In this report, we describe the application of an EASE vector system that is capable of generating stable pools of transfected Chinese hamster ovary cells. These pooled populations of cells produce high quantities of antibody candidates without labor-intensive cloning in a 3-5 week time frame. When an optimal drug candidate has been selected, pools generated with EASE-containing vectors can also be used in subsequent cloning steps to make cell lines with improved expression levels. We demonstrate that EASE increases expression in nonamplified pools in addition to increasing amplification and viability of clonal cell lines generated with the EASE-containing vectors compared with pools and cell lines generated without EASE.
Aldrich
Teri L
TL
Department of Cell Sciences, Amgen, Inc., 51 University Street, Seattle, Washington 98101, USA.
Viaje
Aurora
A
Morris
Arvia E
AE
D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000378:metabolism*; D016466:CHO Cells / Q000378:metabolism*; D003001:Cloning, Molecular / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D005786:Gene Expression Regulation / Q000502:physiology*; D005822:Genetic Vectors / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014162:Transfection / Q000379:methods*
D000906:Antibodies; D011994:Recombinant Proteins