Figure 2

Production of anti-PD-1 chimeric antibody, Boch1D10F1. A Schematic structures of plasmid vectors encoding Boch1D10F1 (pDC62c5-U533-Boch1D10F1, pDC61-Boch1D10F1, and pNC32c-U533-Boch1D10F1). The light chain consisting of a variable region (VL) and a constant region (CL). The heavy chain consisting of a variable region (VH) and a constant region (CH). mDHFR: modified dihydrofolate reductase, UCOE: ubiquitous chromatin opening element (UCOE), npt: neomycin-resistant gene. B Selection of CHO DG44 cell clones producing Boch1D10F1. The production capacity of Boch1D10F1 from each cell clone transfected with pDC62c5-U533-Boch1D10F1 (blue bars), pDC61-Boch1D10F1 (red bars), and pNC32c-U533-Boch1D10F1 (green bars) are shown. The production capacity of the highest producing clones for each vector are shown in the plot. C Expression of Boch1D10F1. Boch1D10F1 was expressed in 2L of shaking culture of a higher-producing cell line. The antibody production (left axis: gray bar) and the density of live and dead cells (right axis: black and white circles) were measured at 3- to 4-day intervals. D Purification of Boch1D10F1. Boch1D10F1 was purified from the supernatants of shaking cultures. Purified protein was confirmed by reducing and nonreducing SDS-PAGE. An uncropped gel image generated during the current study is provided in Additional file 4.