Sustained activation and tumor targeting of NKT cells using a CD1d–anti-HER2–scFv fusion protein induce antitumor effects in mice
J. Clin. Invest. Kathrin Stirnemann, et al. 118:994 doi:10.1172/JCI33249 [Go to this article.]

Figure 1
Design, production, and purification of recombinant CD1d molecules. (A) Design of the genetic fusions of mouse β₂m with the sCD1d and the single chain of the murine anti-HER2 antibody 4D5 (4D5 scFv). DNA fragments were produced by PCR using primers, including sequences for flexible glycine-serine rich linkers (G₁₀S₃ and G₃S₃). A His-tag (His)₆ was added at the C-terminus for Ni-NTA purification. (B) sCD1d and sCD1d–anti-HER2 fusion proteins after production in HEK293EBNA cells. Shown are 10% SDS-PAGE after purification of the proteins by Ni-NTA chromatography (upper panel) and FPLC Sephacryl S100 profiles after loading with αGalCer (lower panels). β₂m-L, leader sequence of β₂m; α1-3, extracellular domains of CD1d; M, molecular weight markers.