广东工业大学学报 ›› 2012, Vol. 29 ›› Issue (3): 87-91.doi: 10.3969/j.issn.1007-7162.2012.03.017
黄明星,叶韵,谭竹钧,韩雅莉
Huang Mingxing, Ye Yun, Tan Zhujun, Han Yali
摘要: 介绍一种改良的用以快速地定性和半定量检测纤溶活性蛋白的纤维平板法.对该方法中的几个关键因素作了详细的研究和探讨.纤维平板中纤维蛋白原和纤溶酶原的浓度对裂解面积不产生任何影响,但是纤维蛋白原的浓度会对裂解圈的观察产生影响,纤维蛋白原浓度低于02 mg/mL时会使背景色过淡而不能清晰地观察到裂解圈.尿激酶浓度在2~250 mU/μL范围时,尿激酶浓度的对数和裂解圈相互垂直的两条直径乘积的对数之间存在明显的线性关系.在本实验设计方案下,尿激酶的最佳使用浓度为10~20 mU/μL.同时,本文还探讨了纤维平板中纤溶酶原的去活条件,当琼脂纤维平板凝固后很难完全去活纤溶酶原,但是,在琼脂纤维蛋白原混合物凝固前于95 ℃持续温浴40 min,则能彻底去活纤维蛋白原中残留的纤溶酶原.
[1] Permin P M. Properties of the fibrinokinasefibrinolysin system[J]. Nature(Lond), 1947(160): 571572.[2] Permin P M. The fibrinolytic activator in animal tissue[J]. Acta Physiologica Scandinavica, 1950(21): 159167.[3] Permin P M. 2 simple methods of determining fibrinolytic enzymes[J]. Acta Physiologica Scandinavica, 1950(20): 388393.[4] Astrup T, Mullertz S. The fibrin plate method for estimating fibrinolytic activity[J]. Archives of Biochemistry and Biophysics, 1952(40): 346351.[5] Lassen M. Heat denaturation of plasminogen in the fibrin plate method[J]. Acta Physiologica Scandinavica, 1953(27): 371376.[6] Walton P L. An improved fibrin plate method for assay of plasminogen activators[J]. Clinica Chimica Acta, 1966(13): 680684.[7] Barta G. Dyed fibrin plate assay of fibrinolysis[J]. Canadian Journal of Physiology and Pharmacology, 1966(44): 233240.[8] Jespersen J, Astrup T. A study of the fibrin plate assay of fibrinolytic agentsoptimal conditions, reproducibility and precision[J]. Haemostasis, 1983(13): 301315.[9] Fossum S, Hoem N O. Urokinase and nonurokinase fibrinolytic activity in proteaseinhibitordeprived plasma, assayed by a fibrin microplate method[J].Immunopharmacology,1996(32):119121.〖ZK)〗[10] Zhao M, Pan Y, Wang Y, et al. Establishment of a sensitive method, thin polyacrylamide fibrin plate assay for detection of fibrinolysis activity in vitro[J]. Chinese Pharmaceutical Journal, 2005(40):302304.[11] Kim SR, Sim JS, Ajjappala H, et al. Expression and largescale production of the biochemically active human tissueplasminogen activator in hairy roots of oriental melon (cucumis melo)[J]. Journal of Bioscience and Bioengineering,2012 (113): 106111.[12] Park J, Yoon S, Kim S, et al. Characterization and fibrinolytic activity of acetobacter sp. FP1 isolated from fermented pine needle extract[J]. Journal of Microbiology and Biotechnology,2012 (22):215219.[13] Simkhada J R, Cho S S, Mander P, et al. Purification, biochemical properties and antithrombotic effect of a novel streptomyces enzyme on carrageenaninduced mice tail thrombosis model[J].Thrombosis Research,2012(129):176182.[14] Uesugi Y, Kawata H, Saito Y, et al. Ultrasoundresponsive thrombus treatment with zincstabilized gelatin nanocomplexes of tissuetype plasminogen activator[J].Journal of Drug Targeting,2012(20):224234. |
No related articles found! |
|