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利用空间排除色谱分离生物高分子,过去都是采用软的亲水性凝胶为柱填料。例如Bio-Gel P(交联聚丙烯酰胺)、Sephadex(交联葡聚糖凝胶)、Sepharose(珠状琼脂糖)、Bio-Gel A(琼脂糖凝胶)等。这些凝胶不能承受较高的柱压,一般只能在0.002-0.02厘米/秒的流速下操作,才能保持凝胶的机械稳定性。这个流速比高效液相色谱所通用的流速约低一个数量级。因此,达到分离平衡的淋洗时间较长,分离周期往往需要几小时。此外,这类软凝胶一般需要较大的进样量方能检出,如对蛋白质的检测就需要毫克级的量。
The use of space exclusion chromatography biopolymer, the past are using soft hydrophilic gel as column packing. Such as Bio-Gel P (cross-linked polyacrylamide), Sephadex (sephadex), Sepharose (beads agarose), Bio-Gel A (agarose gel) and the like. These gels can not withstand higher column pressures and are generally only operated at flow rates of 0.002-0.02 cm / sec in order to maintain the mechanical stability of the gel. This flow rate is about one order of magnitude lower than that common to high performance liquid chromatography. Therefore, to achieve a balanced separation of the leaching longer, the separation cycle often takes a few hours. In addition, soft gels of this type generally require a larger amount of sample to be detected, as is the case with proteins that require milligram quantities.