Use of high-performance size-exclusion chromatography to measure protein molecular weight and hydrodynamic radius. An investigation of the properties of the TSK 3000 SW column.

Use of high-performance size-exclusion chromatography to measure protein molecular weight and hydrodynamic radius. An investigation of the properties of the TSK 3000 SW column.
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使用高性能尺寸排阻色谱法测量蛋白质分子量和流体动力学半径。

DOI:
10.1111/j.1399-3011.1985.tb01021.x
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发表时间:
1985
期刊:
International journal of peptide and protein research
影响因子:
--
通讯作者:
Church,FC
Church,FC
中科院分区:
--
文献类型:
--
作者:
Tarvers,RC;Church,FC

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我们对TSK 3000 SW高性能分子排阻色谱柱进行了一项研究,以确定在何种条件下蛋白质迁移与其已知的流体动力学性质最一致。结果表明:1)在低离子强度下,柱上残留的负电荷确实会引起电荷排斥或电荷保留效应,在去离子水中洗脱时,几种阴离子蛋白质约在空隙体积内; 2)当μ > 0.5时,蛋白质迁移不仅与离子强度无关,而且与蛋白质分子量和流体力学体积一致; 3)小的疏水肽被柱阻滞;和4)非常不对称的蛋白质和其他流体动力学颗粒可能被“末端插入”机制阻滞。
We have conducted a study of the TSK 3000 SW high‐performance size‐exclusion column to define under what conditions proteins would migrate most consistently with their known hydrodynamic properties. Our findings include the following: 1) the residual negative charge of the column does cause charge‐exclusion or charge‐retention effects at low ionic strengths; with elution in deionized water several anionic proteins elute approximately in the void volume; 2) at μ > 0.5, protein migration is not only independent of ionic strength,butconsistent with protein molecular weightandhydrodynamic volume; 3) small hydrophobic peptides are retarded by the column; and 4) very asymmetric proteins and other hydrodynamic particles are likely to be retarded by an “end‐on insertion” mechanism.
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