Figure 5
Recovery using EGFP as a flow-through protein indicator. (A) Ten chromatograms from the 20th, 60th, 100th, 140th, 180th, and 200th MARS column runs using EGFP-spiked plasma were overlain, and an overlay of 10 chromatograms from the 21st, 61st, 101st, 141st, 181st, and 201st MARS column runs using EGFP-only was drawn in (B) using the same pattern. (C) One hundred microliters of MARS buffer A was injected into the MARS column as the blank run. This spectrum at UV488 nm generated 2 peaks at 37 min and 43 min from the buffer components. Thus, the 2 peaks were ignored in subsequent experiments. (D) Equal amounts (5 μg) of plasma (P), unbound fraction (UB), and bound fraction (B) from the 20th, 80th, 100th, 140th, 180th, and 200th runs from the 10 chromatograms were loaded onto an 8-12% SDS-PAGE gel and visualized by silver staining. The bands at 25 kDa in the UB lane ere spiked EGFP; such bands were not detected in any other B lane.