Scott J H, Kelner K L, Pollard H B
Anal Biochem. 1985 Aug 15;149(1):163-5. doi: 10.1016/0003-2697(85)90489-0.
Synexin can be purified to virtual homogeneity by a modification of the conventional chromatofocusing technique. Ampholytes are omitted from the procedure altogether and the protein is eluted by a specific pH step chosen on the basis of the pI of the protein. This modification of the conventional method eliminates the separation of ampholytes from the purified protein, an insurmountable problem in our case, and reduces the cost of purification profoundly.
通过对传统色谱聚焦技术进行改进,可将钙结合蛋白纯化至近乎同质的状态。在该方法中完全省去两性电解质,根据蛋白质的等电点选择特定的pH梯度来洗脱蛋白质。这种对传统方法的改进消除了从纯化蛋白中分离两性电解质的问题(在我们的案例中这是一个无法解决的问题),并大幅降低了纯化成本。