HIGH-PERFORMANCE GEL-PERMEATION CHROMATOGRAPHY OF PROTEINS AND PEPTIDES ON COLUMNS OF TSK-G2000-SW AND TSK-G3000-SW - A VOLATILE SOLVENT GIVING SEPARATION BASED ON CHARGE AND SIZE OF POLYPEPTIDES
HIGH-PERFORMANCE GEL-PERMEATION CHROMATOGRAPHY OF PROTEINS AND PEPTIDES ON COLUMNS OF TSK-G2000-SW AND TSK-G3000-SW - A VOLATILE SOLVENT GIVING SEPARATION BASED ON CHARGE AND SIZE OF POLYPEPTIDES
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DOI：
10.1016/0003-2697(86)90239-3
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发表时间：
1986-05-15
影响因子：
2.9
通讯作者：
SHAW, C
中科院分区：
文献类型：
--
作者：
IRVINE, GB;SHAW, C
三氟乙酸 (0.1% w/v) 是多肽的优良溶剂，具有挥发性，并且对低波长紫外线的吸光度较低。在该溶剂中，在TSK-G2000-SW或TSK-G3000-SW柱上进行层析的多肽被洗脱为尖锐的峰。保留体积不仅取决于分子量，还取决于每个分子的形式电荷数。对于分子量相近的多肽，碱性氨基酸残基比例最高的多肽最早被洗脱。对于TSK-G2000-SW色谱柱，洗脱体积与分子量对数之间线性关系的偏离程度与分子量除以每个分子正电荷数的2/3次方成正比。溶剂中包含 0.25 M 氯化钠增加了发生分离的分子量范围的上限和下限。因此，使用0.1%三氟乙酸作为流动相特别适用于低分子量肽的分离。
Trifluoroacetic acid (0.1% w/v) is an excellent solvent for polypeptides, is volatile, and has a low absorbance of ultraviolet light of low wavelength. Polypeptides subjected to chromatography on columns of TSK-G2000-SW or TSK-G3000-SW in this solvent were eluted as sharp peaks. Retention volume was dependent not only on molecular weight but also on the number of formal charges per molecule. For polypeptides with a similar molecular weight, that with the highest proportion of basic amino acid residues was eluted earliest. For the TSK-G2000-SW column, the degree of deviation from a linear relationship between elution volume and logarithm of molecular weight was directly proportional to the molecular weight to the power 2/3 divided by the number of positive charges per molecule. Inclusion of 0.25 M sodium chloride in the solvent increased both the upper and lower limits of the molecular weight range over which separation occurred. The use of 0.1% trifluoroacetic acid as the mobile phase is thus particularly applicable to the separation of peptides of low molecular weight.