Warner Christopher J A, Dutta Subrata, Foley Alejandro R, Raskatov Jevgenij A
Department of Chemistry, University of California, Santa Cruz.
Department of Chemistry, University of California, Santa Cruz;
J Vis Exp. 2017 Mar 27(121):55482. doi: 10.3791/55482.
Amyloidogenic peptides such as the Alzheimer's disease-implicated Amyloid beta (Aβ), can present a significant challenge when trying to obtain high purity material. Here we present a tailored HPLC purification protocol to produce high-purity amyloid beta 42 (Aβ42) and amyloid beta 40 (Aβ40) peptides. We have found that the combination of commercially available hydrophobic poly(styrene/divinylbenzene) stationary phase, polymer laboratory reverse phase - styrenedivinylbenzene (PLRP-S) under high pH conditions, enables the attainment of high purity (>95%) Aβ42 in a single chromatographic run. The purification is highly reproducible and can be amended to both semi-preparative and analytical conditions depending upon the amount of material wished to be purified. The protocol can also be applied to the Aβ40 peptide with identical success and without the need to alter the method.
诸如与阿尔茨海默病相关的β淀粉样蛋白（Aβ）等淀粉样生成肽，在试图获得高纯度材料时可能会带来重大挑战。在此，我们提出一种定制的高效液相色谱（HPLC）纯化方案，以生产高纯度的β淀粉样蛋白42（Aβ42）和β淀粉样蛋白40（Aβ40）肽。我们发现，市售的疏水性聚（苯乙烯/二乙烯基苯）固定相，即聚合物实验室反相 - 苯乙烯二乙烯基苯（PLRP - S），在高pH条件下，能够在一次色谱运行中获得高纯度（>95%）的Aβ42。该纯化具有高度的可重复性，并且可以根据希望纯化的材料量调整为半制备和分析条件。该方案也可以同样成功地应用于Aβ40肽，而无需改变方法。
