activity gradient
1.Human early pregnncy factor (EPF) was purified fron the pooled serum of 110 pregnant women between 3 and 8 weeds of gestation using ionexchange Chromatography, Affinity Chromatography and linear gradient elution technique·EPF activity of the exract at various stage wasdetected by the erythrocty active rasetle inhibition test (Ea-RIT)·Preghant H3 of the last extractwas proved of EPF activity.
2.Three kinds of CpG ODN 2 006, D19 and 1826 identified optimally activity in human, swine and mice were se lected and synthesized respectively, and so was the control ODN D48 which was no CpG-containing oligodeoxynucleotides. Swine peripheral blood mononuclear cells (PBMC) were prepared by density gradient centrifugation, mouse spleen lymphocy tes by resolving erythrocyte with NH4Cl;
4.In the CM-cellulose ion-exchange chromotography, the volume of MTG was 40mL, on the linear gradient elution method, MTG could be eluted by 300mL, 0~0.3mol/L phosphate buffer, at a flow tate of 0.4 mL/min. the prification fold was 13.6, while the specific activity and the recovery were 25U/mg and 63%, respectively.
5.ResultsThe chomosone number of NFS 60 cell was 39, about 1×10 5 cells/ml to 1×10 7 cells/ml,the dying color showed gradient when the NFS 60 cells was dyed by MTT,the method(4,4) was adopted in the determination of rhG CSF biological activity,the average FL% of potency was 13.560% for single expriment,and the CV of inter assay and intra assay was lower than 10% and 10.109%,respectively.
