Arakawa Tsutomu, Tokunaga Masao, Maruyama Takuya, Shiraki Kentaro
Alliance Protein Laboratories, A Division of KBI Biopharma, 6042 Cornerstone Court West, San Diego, CA 92121, United States.
Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.
Curr Protein Pept Sci. 2019;20(1):28-33. doi: 10.2174/1389203718666171117105132.
MEP (mercapto-ethyl-pyridine) HyperCel is one of the hydrophobic charge induction chromatography (HCIC) resins. Under normal operation, proteins are bound to the MEP resin at neutral pH, at which MEP is not charged, mostly via hydrophobic interaction. MEP has a pyridine group, whose pK is 4.8, and hence is positively charged at acidic pH range. Based on the binding mechanism (i.e., hydrophobic interaction) and the induced positive charge at acidic pH, there may be two ways to elute the bound proteins. One way is to bring the pH down to protonate both MEP resin and the bound protein, leading to charge repulsion and thereby elution. Another way is to use hydrophobic interaction modifiers, which are often used in hydrophobic interaction chromatography, to reduce hydrophobic interaction. Here, we summarize such two possible elution approaches.
巯基乙基吡啶（MEP）HyperCel是一种疏水电荷诱导色谱（HCIC）树脂。在正常操作下，蛋白质在中性pH值时与MEP树脂结合，此时MEP不带电荷，主要通过疏水相互作用结合。MEP有一个吡啶基团，其pK为4.8，因此在酸性pH范围内带正电荷。基于结合机制（即疏水相互作用）以及在酸性pH下诱导产生的正电荷，可能有两种洗脱结合蛋白质的方法。一种方法是降低pH值使MEP树脂和结合的蛋白质都质子化，导致电荷排斥从而实现洗脱。另一种方法是使用疏水相互作用调节剂，这在疏水相互作用色谱中经常使用，以减少疏水相互作用。在此，我们总结这两种可能的洗脱方法。
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