Related Researching About F?? In Fibrinogen

F?? is a transglutaminase,it covalently cross-links the glutamine and lysine residues of forming fibrin strands and thus increases clot strength and stability.The five main measurements of F?? activity are clot solubility test,ammonia release assay,Amine incorporation assay,ELISA and Fluorescent Labeling.By compared the results of clot solubility test and ammonia release assay,we determined the suitable method for freeze-dried fibrinogen F?? activity analysis.We reduced the cost of daily analysis by using freeze-dried pig standard plasma,and we ensured the quality of analysis by control the quality of freeze-dried pig standard plasma and F??-deficient fibrinogen.In chapter 1,we briefly introduced the background of this study.First,we introduced the structure and function of F??.Then,we summarized the situation of domestic Fibrin Sealant.In addition,we summarized the testing method of F??.In chapter 2,we researched freeze-dried pig standard plasma production process and quality standard.We used mannitol and HEPES as protective agent of freeze-dried pig standard plasma.Freeze-dried pig standard plasma quality standard has five indicators,solute time,sterility,water,loading quantity precision and content uniformity.Three batches freeze-dried pig plasma were all meet the requirements of quality standard,their F?? activity are 100.29%,100.37% and 100.88%?In chapter 3,we established F??-deficient fibrinogen quality standard.The quality standard has three indicators,appearance,clotsolubility examination and fibrinogen content.The detection limit of clot solubility examination was 0.5-1.0%.We choose Von Clauss method for content determination of fibrinogen in F??-deficient fibrinogen.It's a highly significant double logarithmic linear relationship between the fibrinogen concentration and coagulation time(r =0.9988).The linear range of fibrinogen was 104-624 mg/L.The reproducibility,precision,and accuracy of the method were good.In chapter 4,we choose modified clot solubility test for activity determination of F?? in freeze-dried pig fibrinogen.And we performed the method validation for it.The specificity,specificity,reproducibility,and stability of the method were good.It can be used to effectively control the quality of the product.The physiological saline containing 0.4% human blood albumin was used to dilute the sample,making F?? uniform distribution in solution.We investigate the effect of the thrombin.We measured the freeze-dried fibrinogen from different sources.The data from modified clot solubility test were more close to the real activity.In chapter 5,we used clot solubility test,modified clot solubility test and ammonia release assay for the determination of F?? activity in Plasma(human,pig and mixture).For F?? activity in human Plasma,the determination results of ammonia release assay group and modified clot solubility test group are 0.998±0.189U/ml,1.086±0.462U/ml.There were no significant differences between the two groups(P=0.221).For F?? activity in pig Plasma,the determination results of ammonia release assay group and modified clot solubility test group are 0.955±0.194U/ml,1.043±0.443U/ml.There were no significant differences between the two groups(P=0.115).There were no significant differences of F?? activity between pig Plasma and human Plasma(P=0.355).The results of ammonia release assay and modified clot solubility test are very similar.In chapter 6,we summarized the whole project and looked into thefuture.