Comparison of the separation of proteins of wide‐ranging molecular weight via trilobal polypropylene capillary‐channeled polymer fiber, commercial superficiously porous, and commercial size exclusion columns
Comparison of the separation of proteins of wide‐ranging molecular weight via trilobal polypropylene capillary‐channeled polymer fiber, commercial superficiously porous, and commercial size exclusion columns
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通过三叶聚丙烯毛细管通道聚合物纤维、商用表面多孔柱和商用尺寸排阻柱分离宽范围分子量蛋白质的比较
DOI：
10.1002/jssc.202100891
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发表时间：
2022
影响因子：
3.1
通讯作者：
Marcus, Richard Kenneth
中科院分区：
文献类型：
--
作者：
Huang, Sisi;McClain, Ray T.;Marcus, Richard Kenneth
反相和尺寸排阻色谱法通常用于蛋白质分离，尽管它们基于截然不同的原理。反相方法在多孔支持物上产生基于疏水性的（松散术语）蛋白质分离，但基于传质限制，倾向于限于具有适度分子量的蛋白质。或者，尺寸排阻在基于熵而非熵的过程的较高质量蛋白质的分离中提供互补益处，但倾向于产生有限的峰容量。在这项研究中，微孔柱填充了一种新的三聚氰胺聚丙烯毛细管通道聚合物纤维，用于分离的多肽和蛋白质的分子量范围从1.4到660 kDa的反相模式。优化色谱参数，包括梯度时间、流速和移动的相中的三氟乙酸浓度，以最大限度地提高分离度和通量。优化后，将triethylene纤维色谱柱的性能与两种商业来源的色谱柱（表面多孔C4衍生化硅胶和尺寸排阻色谱柱）进行了比较，这两种色谱柱均专门用于蛋白质分离，并根据制造商规定的条件进行操作。与商业色谱柱相比，纤维色谱柱在整个系列中具有更好的分离性能，成本更低，分离时间更短。
Reversed phase and size‐exclusion chromatography methods are commonly used for protein separations, although they are based on distinctly different principles. Reversed phase methods yield hydrophobicity‐based (loosely‐termed) separation of proteins on porous supports, but tend to be limited to proteins with modest molecular weights based on mass transfer limitations. Alternatively, size‐exclusion provides complementary benefits in the separation of higher mass proteins based on entropic, not enthalpic, processes, but tend to yield limited peak capacities. In this study, microbore columns packed with a novel trilobal polypropylene capillary‐channeled polymer fiber were used in a reversed phase modality for the separation of polypeptides and proteins of molecular weights ranging from 1.4 to 660 kDa. Chromatographic parameters including gradient times, flow rates, and trifluoroacetic acid concentrations in the mobile phase were optimized to maximize resolution and throughput. Following optimization, the performance of the trilobal fiber column was compared to two commercial‐sourced columns, a superficially porous C4‐derivatized silica and size exclusion, both of which are sold specifically for protein separations and operated according to the manufacturer‐specified conditions. In comparison to the commercial columns, the fiber‐based column yielded better separation performance across the entirety of the suite, at much lower cost and shorter separation times.