Aida Y, Pabst M J
Dental Research Center, University of Tennessee, Memphis 38163.
J Immunol Methods. 1990 Sep 14;132(2):191-5. doi: 10.1016/0022-1759(90)90029-u.
Endotoxin contamination of protein solutions was reduced by a phase separation technique using the detergent, Triton X-114. Protein solutions containing endotoxin were treated with Triton X-114 on ice. The solution was then warmed to 37 degrees C, whereupon two phases formed. The Triton X-114 phase, containing the endotoxin, was precipitated by centrifugation. The first cycle of phase separation produced a 1000-fold reduction of endotoxin from contaminated preparations of cytochrome c, catalase and albumin. Complete removal of endotoxin could be achieved by further cycles of phase separation. Each cycle of phase separation resulted in only a 2% loss of protein, and could be completed within 15 min. The small amount of detergent (0.018%) that persisted in protein solution could be removed by gel filtration or absorption. Proteins treated by this procedure retained normal functions. This phase separation technique provides a rapid and gentle method for removing endotoxin from protein solutions.
采用去污剂Triton X-114的相分离技术降低了蛋白质溶液中的内毒素污染。将含有内毒素的蛋白质溶液在冰上用Triton X-114处理。然后将溶液加热至37℃，此时形成两相。含有内毒素的Triton X-114相通过离心沉淀。相分离的第一个循环使细胞色素c、过氧化氢酶和白蛋白污染制剂中的内毒素减少了1000倍。通过进一步的相分离循环可以实现内毒素的完全去除。每个相分离循环仅导致2%的蛋白质损失，并且可以在15分钟内完成。蛋白质溶液中残留的少量去污剂（0.018%）可通过凝胶过滤或吸附去除。用该方法处理的蛋白质保留了正常功能。这种相分离技术为从蛋白质溶液中去除内毒素提供了一种快速且温和的方法。
