Wu L H, Tsai C M, Frasch C E
Anal Biochem. 1987 Feb 1;160(2):281-9. doi: 10.1016/0003-2697(87)90048-0.
A new gel filtration method was developed for purification of R-type lipopolysaccharides (lipooligosaccharides) from some nonenteric gram-negative bacteria, including Neisseria meningitidis, Haemophilus influenzae, and Bordetella pertussis. These wild-type lipooligosaccharides are poorly extractable by the phenol-chloroform-ether extraction method of C. Galanos, O. Luderitz, and O. Westphal [1969) Eur. J. Biochem. 9, 245-249) and therefore a new procedure was developed for their isolation. The lipooligosaccharides (LOS) were first extracted by hot phenol-water, treated with RNase, then disaggregated in deoxycholic acid, and purified by gel filtration on Sephadex G-75. By comparison the conventional hot phenol-water purification method using repeated ultracentrifugations yielded less LOS. The yield of LOS by gel filtration was 30 to 108% higher and the purity was better.
开发了一种新的凝胶过滤方法，用于从一些非肠道革兰氏阴性细菌中纯化R型脂多糖（脂寡糖），这些细菌包括脑膜炎奈瑟菌、流感嗜血杆菌和百日咳博德特氏菌。这些野生型脂寡糖很难通过C. Galanos、O. Luderitz和O. Westphal [1969年，《欧洲生物化学杂志》9, 245 - 249页] 的酚 - 氯仿 - 乙醚提取法提取，因此开发了一种新的分离方法。脂寡糖（LOS）首先通过热酚 - 水提取，用核糖核酸酶处理，然后在脱氧胆酸中解聚，并通过Sephadex G - 75凝胶过滤进行纯化。相比之下，使用反复超速离心的传统热酚 - 水纯化方法得到的LOS较少。通过凝胶过滤得到的LOS产量高30%至108%，纯度也更好。
