Somlyo B, Kovats E, Keler T, Nowotny A
University of Pennsylvania, Center for Oral Health Research, Philadelphia 19104.
J Chromatogr. 1990 Feb 23;525(2):329-38. doi: 10.1016/s0378-4347(00)83409-x.
A new, fast and highly reproducible column liquid chromatographic method was elaborated for the analysis and small-scale preparative isolation of endotoxin from Serratia marcescens Bizio (ATCC No. 264). This procedure detects contaminants of such preparations with high sensitivity and it is capable of separating them from endotoxic components. Extensive heterogeneity of both 5% trichloroacetic acid and phenol-water-extracted endotoxin preparations was recorded. Heterogeneity among the endotoxic components of purified preparations could also be detected by this method. Measurements of biological activities, such as Limulus amoebocyte lysate activation, lymphoblastogenesis (mitogenicity) and tumor necrosis factor (TNF) liberation were carried out on the chromatographically separated fractions. During these studies, non-toxic but in vitro TNF-generating components of crude endotoxin extracts were also detected.
建立了一种新的、快速且高度可重复的柱液相色谱方法，用于分析和小规模制备分离粘质沙雷氏菌Bizio（ATCC编号264）中的内毒素。该方法能高灵敏度检测此类制剂中的污染物，并能将其与内毒素成分分离。记录了5%三氯乙酸和酚水提取的内毒素制剂的广泛异质性。该方法还能检测纯化制剂内毒素成分之间的异质性。对色谱分离的组分进行了生物活性测定，如鲎试剂激活、淋巴细胞生成（促有丝分裂性）和肿瘤坏死因子（TNF）释放。在这些研究中，还检测到了粗制内毒素提取物中无毒但在体外能产生TNF的成分。
