Reversed-phase high-performance liquid chromatographic purification and identification of goat HSV-1 specific transfer factor active components
QI Hai-Yan, WAN Zhi-Fang and SU Cheng-Zhi Department of Biochemistry
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.
【CateGory Index】： R392.1