Preparative reversed-phase high-performance liquid chromatography collection efficiency for an antimicrobial peptide on columns of varying diameters (1 mm to 9.4 mm ID)
Preparative reversed-phase high-performance liquid chromatography collection efficiency for an antimicrobial peptide on columns of varying diameters (1 mm to 9.4 mm ID)
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DOI：
10.1016/j.chroma.2006.11.052
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发表时间：
2007-01-26
影响因子：
4.1
通讯作者：
Hodges, Robert S.
中科院分区：
文献类型：
--
作者：
Chen, Yuxin;Mant, Colin T.;Hodges, Robert S.
本研究考察了反相高效液相色谱（RP-HPLC）柱直径（1 mm至9.4 mm I.D.）一步缓慢梯度制备纯化的26个残基的合成抗菌肽。当放在一起时，半制备柱（9.4mm I.D.）在宽的样品负载范围（0.75-200 mg）内提供了纯化产物的最高产率（从亲水性和疏水性杂质的平均回收率为90.7%）。直径较小的色谱柱，如窄孔色谱柱（150 x 2.1 mm I.D.）和微孔柱（150 x 1.0 mm内径），可用于纯化肽，具有纯化产物的合理回收，但可应用于柱的粗肽的范围有限。此外，较小直径的柱需要更广泛的馏分分析，以定位具有相同负载的较大直径的柱的纯产物的馏分。我们的研究结果表明，一步缓慢梯度制备协议作为一个通用的方法纯化合成肽的优秀潜力。(c)2006 Elsevier B. V.保留所有权利。
The present study examines the effect of reversed-phase high-performance liquid chromatography (RP-HPLC) column diameter (1 mm to 9.4 mm I.D.) on the one-step slow gradient preparative purification of a 26-residue synthetic antimicrobial peptide. When taken together, the semi-preparative column (9.4 mm I.D.) provided the highest yields of purified product (an average of 90.7% recovery from hydrophilic and hydrophobic impurities) over a wide range of sample load (0.75-200 mg). Columns with smaller diameters, such as narrowbore columns (150 x 2.1 mm I.D.) and microbore columns (150 x 1.0 mm I.D.), can be employed to purify peptides with reasonable recovery of purified product but the range of the crude peptide that can be applied to the column is limited. In addition, the smaller diameter columns require more extensive fraction analysis to locate the fractions of pure product than the larger diameter column with the same load. Our results show the excellent potential of the one-step slow gradient preparative protocol as a universal method for purification of synthetic peptides. (c) 2006 Elsevier B.V. All rights reserved.