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. 2016 Dec 15;6:39221. doi: 10.1038/srep39221

Figure 2. Affinity chromatography of fluorescence-labeled rFab containing E tag on the unified capillary.

Figure 2

The unified capillary (50 μm i.d., 48.5 cm in length) was equilibrated with PBS-Tw at 2.0 psi (a linear flow rate of 15 cm/min) for 5 min. The fluorescence-labeled rFab at 100 nM in PBS-Tw was applied at 0.8 psi (6.0 cm/min) for 15 min (1.75 μL, 175 fmol of rFab) on the capillary bearing anti-E tag antibody. The column was washed at 2.0 psi for 2 min each with PBS-Tw containing 0.5 M NaCl, and with 20 mM Tris-HCl (pH 7.4) containing 0.1% (w/v) Tween 20. The bound material was eluted with 100 mM phosphoric acid at 2.0 psi for 5 min (green solid line). Exactly the same procedure was carried out on the capillary before immobilizing the anti-E tag antibody, i.e., only with the immobilized streptavidin (blue dotted line). The pressure (psi) was recorded with the black line. RFU, relative fluorescence unit.