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Equilibration, sample, and washings with Buffers A and B were carried out as indicated in Section 4.1, Section 4.2.1, and Section 4.3. Next, 10 CV of linear gradient 5 mM to 25 mM CaCl2 (in Buffer B) and 5 CV stepwise Buffer B + 25 mM CaCl2 were carried out. They were then washed with … Read more…
The absence of the peak 500 in Figure 4a and the similarity of the protein profile lane 65A of Figure 4a (purification on Q resin) and lane “500” of Figure 4d,f (purification on ANX) indicate that FVIII eluted due to increase of the ionic strength. Furthermore, work with highly concentrated Ca2+ solutions is not recommended … Read more…
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