RAPID PROTEIN SEPARATION AND DIFFUSION-COEFFICIENT MEASUREMENT BY FRIT INLET FLOW FIELD-FLOW FRACTIONATION
RAPID PROTEIN SEPARATION AND DIFFUSION-COEFFICIENT MEASUREMENT BY FRIT INLET FLOW FIELD-FLOW FRACTIONATION
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DOI：
10.1002/pro.5560020917
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发表时间：
1993-09-01
期刊：
影响因子：
8
通讯作者：
GIDDINGS, JC
中科院分区：
文献类型：
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作者：
LIU, MK;LI, P;GIDDINGS, JC
在这项研究中，三个流场-流动分馏(Flow Fff)通道被用于分离蛋白质并同时测量它们的平移扩散系数，D.一个通道具有传统的进样通道，而另外两个通道采用熔块进样设计，允许更方便和快速地进样。本文从理论上描述了保留时间对D的依赖关系，并从实验上验证了流动FFF纯化的蛋白质峰与D的关系。还研究了影响频带展宽、分辨率和优化的因素。分离蛋白质的时间范围为4-20分钟。在每次需要2分钟的多次运行中实现部分解析。根据保留时间计算了15种蛋白质的D值。其中包括两种通常无法测量的蛋白质二聚体(牛血清白蛋白和丙种球蛋白)。将三个渠道的D值相互比较，并与文献数据进行比较。找到了合理的一致性(在3-4%范围内)。高速重复运行可用于在短至1分钟的时间间隔内获取多个D值。
In this study three flow field-flow fractionation (flow FFF) channels are utilized for the separation of proteins and for the simultaneous measurement of their translational diffusion coefficients, D. One channel has a traditional sample inlet, whereas the other two incorporate a frit inlet design that permits more convenient and rapid sample introduction. The dependence of retention time on D, which leads to differential elution and the opportunity to measure D for protein peaks purified by the flow FFF process, is described theoretically and examined experimentally. Factors affecting band broadening, resolution, and optimization are also examined. The separation of proteins is achieved in the time range 4-20 min. Partial resolution is achieved in multiple runs requiring 2 min each. Values of D calculated from retention times are reported for 15 proteins. These include two protein dimers (bovine serum albumin and gamma-globulin) not ordinarily accessible to measurement. The D values from the three channels are compared with one another and with literature data. Reasonable consistency (within 3-4%) is found. High-speed repetitive runs can be used to acquire multiple values of D in time intervals as short as 1 min.