The Establishment And Rudiment Application Of Two Methods Of ELISA For N-V Proteinase

N-V proteinase is a fibrinolysin from a nature marine life in our country, it possess fibrinolytic activity and have the effect of thrombolysis. It is a single chain molecule and molecule weight is 31000 u, PI is 4.5, the optimization PH is 7.8, the optimization temperature is 45℃.The mechanism of N-V proteinase is similar to fibrinolysin in human, it can dissolve fibrin directly. N-V proteinase possess the fibrinobytic activity in vitro, it obviously decrease the content of Fg and D-Dimer, and lessen ELT, while have not influence the APTT and PT. The experiment of thrombolysis intends that N-V proteinase is superior to UK in the thrombolysis. Absolutely, N-V proteinase has a good practice value.N-V proteinase is a kind of aminopeptidase, the aminopeptidase can hydrolyze amino acids from the NH2-terminal and then hydrolyze protein. So when we prepared the polyclonal antibody, we can not inject the N-V proteinase to the muscle of animals indirectly. So we deal with it by the application of adjuvant to lessen the hydrolization activity, then we get the polyclonal antibody of N-V proteinase. The polyclonal antibody of N-V proteinase valence is 1:64 by the double immunodiffusion electrophoresis, and the antibody has a strong immunologic specificity by the fuse sediment arc.In order to isolate IgG, we employ the method of ammonium sulphate precipitation and phenyl-superose FF chromatography.Sodium sulphate polyacrylamide gel electrophoresis indicates that we obtain relative pure IgG .Densitometry scanning indicated that the purity was 95%. We exploit the...