Josić D, Baumann H, Reutter W
Anal Biochem. 1984 Nov 1;142(2):473-9. doi: 10.1016/0003-2697(84)90492-5.
When size-exclusion HPLC (SE-HPLC) is applied for the separation of hydrophilic and hydrophobic proteins, numerous problems can be encountered, which may present considerable difficulties. A major source of such complications is interaction between column packing and sample, especially on the so-called "Diol" and "Polyol" columns. In many cases interaction can be reduced only by adding detergents. Calibration proteins and hydrophobic membrane proteins of liver are separated by SE-HPLC. The influence of detergents on association and dissociation of protein subunits and protein configuration is shown. These factors can affect the elution volume during chromatography. Furthermore it is shown that a direct comparison can be drawn between the protein separation by SE-HPLC on the one hand and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis on the other hand. As an example the separation of the delta-subunit of the acetylcholine receptor was shown under reducing and nonreducing conditions.
当采用尺寸排阻高效液相色谱法（SE-HPLC）分离亲水性和疏水性蛋白质时，可能会遇到许多问题，这些问题可能带来相当大的困难。此类复杂情况的一个主要来源是柱填料与样品之间的相互作用，尤其是在所谓的“二醇”柱和“多元醇”柱上。在许多情况下，只有通过添加去污剂才能减少相互作用。通过SE-HPLC分离校准蛋白和肝脏的疏水膜蛋白。展示了去污剂对蛋白质亚基的缔合和解离以及蛋白质构象的影响。这些因素会影响色谱过程中的洗脱体积。此外，结果表明，一方面通过SE-HPLC进行的蛋白质分离与另一方面通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进行的蛋白质分离之间可以进行直接比较。例如，展示了在还原和非还原条件下乙酰胆碱受体δ亚基的分离情况。
