Allen R G, Barrett J T, Campbell B J
Appl Microbiol. 1971 Jan;21(1):53-60. doi: 10.1128/am.21.1.53-60.1971.
A lipoprotein inhibitor of Newcastle disease virus was obtained from chicken lung tissue by means of dilute alkaline extraction procedures. The inhibitor was further purified by ammonium sulfate fractionation, isoelectric precipitation, and density gradient centrifugation. The purified lipoprotein inhibited active Newcastle disease virus hemagglutination at a concentration of 2.0 mug/ml which represented a 30-fold purification over the original extract. Infection of chicken embryo fibroblasts by Newcastle disease virus was also inhibited by the purified lipoprotein, the degree of inhibition depending upon the inhibitor-to-virus ratio. Chemical analysis of the purified inhibitor provided a composition of 72% lipid, 26% protein, and 3% carbohydrate, although some compositional variation was observed from one preparation to another. The chloroform-soluble lipids were shown to contain 40 to 50% phospholipid and 10 to 20% cholesterol; of the fatty acids recovered from the saponified lipoprotein, 39% was palmitic, 22% oleic, and 17% stearic. Careful analyses of large quantities of the inhibitor revealed a small (0.84%) but significant content of sialic acid. Removal of sialic acid from the lipoprotein by means of digestion with neuraminidase produced a sharp diminution in inhibitory properties. A delipidized form of the inhibitor was obtained by ether extraction, and this material produced a single broad band of precipitate in gel immunodiffusion tests.
通过稀碱提取法从鸡肺组织中获得了新城疫病毒的脂蛋白抑制剂。该抑制剂通过硫酸铵分级分离、等电沉淀和密度梯度离心进一步纯化。纯化后的脂蛋白在浓度为2.0微克/毫升时可抑制活性新城疫病毒的血凝反应,相较于原始提取物,纯化倍数达30倍。纯化后的脂蛋白也能抑制新城疫病毒对鸡胚成纤维细胞的感染,抑制程度取决于抑制剂与病毒的比例。对纯化后的抑制剂进行化学分析,其组成为72%的脂质、26%的蛋白质和3%的碳水化合物,不过不同批次的制剂在成分上存在一定差异。氯仿可溶脂质显示含有40%至50%的磷脂和10%至20%的胆固醇;从皂化脂蛋白中回收的脂肪酸,39%为棕榈酸,22%为油酸,17%为硬脂酸。对大量抑制剂进行仔细分析发现含有少量(0.84%)但显著的唾液酸。用神经氨酸酶消化脂蛋白去除唾液酸后,其抑制特性急剧下降。通过乙醚提取获得了抑制剂的脱脂形式,该物质在凝胶免疫扩散试验中产生单一的宽沉淀带。