摘要
作者应用反相高效液相色谱(RP-HPLC)C_(18)柱,对亲和色谱分离的Ⅰ型单疱病毒(HSV-1)特异转移因子(TF_(HSV-1)活性组分进行纯化;分别收集RP-HPLC分离的各组分,用^(51)Cr标记的白细胞粘附抑制试验逐一检测其TF活性;再对其中具有TF活性的组分进行等电聚焦电泳,分析其纯度和等电点.结果显示,RP-HPLC分离的TF第2组分能诱导正常小鼠脾细胞产生对HSV-1的特异粘附抑制效应,具有TF_(HSV-1)抗原特异活性的特征,且此活性与TF_(HSV-1)粗提物产生的活性效应比较,提高约10000倍;该组分经高敏感银染色,在等电聚焦电泳呈现一条电泳区带,其等电点为pH4.48.这些结果表明,经RP-HPLC纯化已得到TF_(HSV-1)的单一活性成分.
HSV-1 specific transfer factor active components (TFC) separated by affinity chromatography were further purified with reversed-phase high-performance liquid chromatography (RP-HPLC). The biological activities of isolated HPLC fractions were evaluated by 51Cr labeled leukocyte adherence inhibition (51Cr LAI) assay, and the purity and isoelectric point (PI) of the fraction responsible for the HSV-1 specific transfer factor (TFHSV-1) activity were then determined by an isoelectric focusing (IEF) electrophoresis in thin-layer polyacrylamide gel.As a result, only the second fraction of TFC eluted from RP-HPLC column produced obvious LAI effect on HSV-1 antigen and had the antigen-specific activity of TFHSV-1. The active moiety showed only a single band on the IEF gel by high-sensitivity silver staining and its PI was pH 4.48. The data presented here demonstrated that a high purified transfer factor active component specific for HSV-1 (PTFC) was obtained by further purification of RP-HPLC.
出处
《第四军医大学学报》
1992年第2期110-112,共3页
Journal of the Fourth Military Medical University
关键词
转移因子
人疱疹病毒
高效液相色谱
transfer lactor
human herpes virus
chromatography
high-performance liquid phase
