Supplementary Figure 1. The impact of elution cycles and elution volume (800 µL – black, 400 µL blue, and 200 µL – red). Supplementary Figure 2. Determination of optimal elution cycles. Protein content was monitored after each elution cycle (800 µL). Non-eluted fraction bound (red) was determined by dividing the measured amount of protein in the eluted well by the difference in protein content of the sample well after sample binding. Total yield (black) is a measurement of the total

Supplementary Figure 1. The impact of elution cycles and elution volume (800 µL – black, 400 µL blue, and 200 µL – red).

Supplementary Figure 2. Determination of optimal elution cycles. Protein content was monitored after each elution cycle (800 µL). Non-eluted fraction bound (red) was determined by dividing the measured amount of protein in the eluted well by the difference in protein content of the sample well after sample binding. Total yield (black) is a measurement of the total protein initially loaded in the sample well (0.79 mg GusA).

Supplementary Figure 3. The effect of protein size on expected maximum protein elution. Minimum Stokes radius was calculated for GusA (4.3 nm), PaS (2.6 nm), and GFP (2.0 nm).

Supplementary Figure 4. Profile of recovered GFP in total yield and recovered mass as a function of initial sample available using FF. The total yield of the purification (black) ranged from 97% to 29%. The total protein recovered (red) ranged from 0.21 to 3.1 mg.