Transient Production of scFv-Fc Fusion Proteins in Mammalian Cells

Single chain Fv (scFv) antibody fragments fused to the human immunoglobulin G (IgG) Fc moiety obtain IgG-like properties, but nevertheless they are still encoded by a single gene. As transient production of scFv-Fc proteins in mammalians can easily achieve milligram amounts, this antibody format is particularly suitable for many research applications. Here, we describe two protocols for the transient secretory production of scFv-Fc antibodies in transformed adherent human embryonic kidney cells HEK293T and suspension HEK293-6E cells. More than 10 �g/mL yield of scFv-Fc protein can be obtained every day for one to two weeks by transient production in HEK293T cells, whereas soluble HEK293-6E cells produce more than 100 �g/mL scFv-Fc protein within 5 days under serum-free conditions.