Dynamic control of protein conformation transition in chromatographic separation based on hydrophobic interactions: molecular dynamics simulation.

摘要: Abstract Conformational transitions of a protein in hydrophobic interaction based chromatography, including chromatography (HIC) and reversed-phase liquid (RPLC), their impact on the separation process performance were probed by molecular dynamics simulation 46-bead β-barrel coarse-grained model confined pore, which represents porous adsorbent. The transition adsorbed from native conformation to an unfolded one occurred as result strong interactions with pore surface, reduced formation aggregates. conformational was also displayed once elution buffer characterized weaker hydrophobicity introduced strip surface. discharged proteins that underwent prone aggregation; thus, unsatisfactory yield obtained. An orthogonal experiment revealed addition strengths protein–protein protein–adsorbent interactions, time required reduce above-mentioned determined HIC RPLC. Stepwise elution, sequential reduction between adsorbent, presented dynamic strategy for tuning favor aggregates during process. obtained this operation higher than steady-state elution. study qualitatively reproduced experimental observations provided insight would be helpful designing optimizing RPLC proteins.