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Protein was eluted with 4CV of 60% buffer B and immediately diluted 10-fold with Buffer A prior to concentration and 3 rounds of buffer exchange to remove excess imidazole by centrifugation at 3095xg at 4 °C in 100 kDa cutoff concentrators (Amicon)

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3b) was selected based on the number and strength of H-bond interactions (H-bond distance and angle), number of hydrophobic interactions, and buried surface area