高纯度人巨细胞病毒包膜抗原的制备。

Preparation of highly purified human cytomegalovirus envelope antigen.

作者信息

Hudecz F, Gonczol E, Plotkin S A

出版信息

Vaccine. 1985 Sep;3(3):300-4. doi: 10.1016/s0264-410x(85)90119-7.

Abstract

A human cytomegalovirus (HCMV) envelope preparation was highly purified by immunoaffinity column chromatography using an anti-cellular-IgG column. The purified envelope induced high titre antibodies to HCMV in guinea-pigs. Analysis of the guinea-pig immune sera by RIA and immunofluorescence (IF) showed that this envelope preparation, unlike its unpurified counterpart, did not induce antibody to cellular contaminants. Dot-blot assay revealed viral proteins in the flow-through fraction and cellular proteins in the bound fractions. Results of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of flow-through and bound fractions suggest that a number of proteins previously identified as virus-specific may, in fact, reflect cellular contamination of the envelope preparation, or crossreactivity of some virus-specific proteins with cellular proteins.

摘要

用人巨细胞病毒（HCMV）包膜制剂通过使用抗细胞IgG柱的免疫亲和柱色谱法进行高度纯化。纯化后的包膜在豚鼠体内诱导出高滴度的抗HCMV抗体。通过放射免疫分析（RIA）和免疫荧光（IF）对豚鼠免疫血清进行分析表明，与未纯化的对应物不同，这种包膜制剂不会诱导针对细胞污染物的抗体。斑点印迹分析显示，流出级分中有病毒蛋白，结合级分中有细胞蛋白。对流出级分和结合级分进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析的结果表明，许多先前被鉴定为病毒特异性的蛋白质实际上可能反映了包膜制剂的细胞污染，或者某些病毒特异性蛋白质与细胞蛋白质的交叉反应性。