Establishment of high concentration Refolding of unstable protein

早些时候，我们正式建立了一个有效的蛋白质重折叠程序梯度去除增溶剂，如尿素。然而，这种方法对不稳定的蛋白质不太有效。我们在这里开发了一个很好的方法来添加蛋白质稳定剂，以便在增溶剂的浓度下获得合理量的折叠蛋白质，其中不稳定的蛋白质不会形成聚集体。我们研究了许多稳定剂，发现60%的浓缩（2.5毫克/毫升）不稳定的蛋白质可以使用40%的甘油作为最好的稳定剂重折叠。该方法可广泛应用于不稳定蛋白质的复性。

Earlier, we formally established an effective refolding procedure for a protein by gradient removal of a solubilizer such as urea. However, this procedure was less effective for unstable proteins. We developed here an excellent method to add protein stabilizer so as to get reasonable amounts of folded protein under the concentration of solubilizer where the unstable protein does not form aggregate. We examined many stabilizers and found that 60% of a concentrated(2.5 mg/ml) unstable protein can be refolded using 40% glycerol as the best stabilizer. This procedure can be widely applicable for the refolding of unstable proteins.