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学科主题: 临床医学
题名:
Production of recombinant human microplasminogen and pilot study in inducing posterior vitreous detachment
作者: Qu, JF; Li, XX; Huang, BR
关键词: Pichia pastoris ; posterior vitreous detachment ; recombinant human microplasminogen
刊名: CURRENT EYE RESEARCH
发表日期: 2005-10-01
DOI: 10.1080/02713680591006282
卷: 30, 期:10, 页:881-889
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Ophthalmology
研究领域[WOS]: Ophthalmology
关键词[WOS]: PLASMIN-ASSISTED VITRECTOMY ; DIFFERENT MOLECULAR FORMS ; MACULAR HOLES ; VITREORETINAL INTERFACE ; TRACTION SYNDROME ; ACTIVE-SITE ; STREPTOKINASE ; ACTIVATION ; EXPRESSION ; UROKINASE
英文摘要:

Purpose : To realize the high production of recombinant human microplasminogen (r-mPlg) with Pichia pastoris and demonstrate the efficacy of r-mPlg in inducing posterior vitreous detachment (PVD). Methods : Recombinant plasmid pAO815-3mPlg was constructed and transformed into SMD1168 cells. Positive recombinant clones were selected with MD plate and cultured in BMG medium, then induced in BMM medium. A protein band corresponding to mPlg with molecular mass of 29 kDa was revealed in SDS-PAGE and confirmed by Western blot. Anion-exchange chromatography and plasminogen activity assay kit were used to obtain purified r-mPlg with biological activity. Twenty eyes of freshly slaughtered pigs were divided into 4 groups, 5 eyes in each group. Group 1 served as normal control. Intravitreal injection of 0.1 ml BSS, 1000 IU/0.1 ml recombinant streptokinase (r-SK) and 1000 IU/0.1 ml r-SK plus 3 U/0.1 ml r-mPlg was done respectively to groups 2, 3, and 4. After incubation at 37 degrees C for 60 min, all eyes were processed for light microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Results : r-mPlg, which has potential fibrinolytic activity, was successfully obtained with yield of 30 mg/L and purity of 97%. PVD was demonstrated by SEM in group 4 but not in other three groups. The retina and the inner limiting membrane (ILM) were well preserved in all eyes. Conclusion : r-mPlg, which has potential fibrinolytic activity, can be produced through Pichia pastoris expression system. Three U of r-mPlg combined with 1000 IU r-SK was effective in producing PVD without damaging the retina.

语种: 英语
WOS记录号: WOS:000233090300009
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/58035
Appears in Collections:北京大学第二临床医学院_期刊论文

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作者单位: 1.Peking Univ, Peoples Hosp, Beijing 100871, Peoples R China
2.Peking Union Med Coll, Beijing, Peoples R China

Recommended Citation:
Qu, JF,Li, XX,Huang, BR. Production of recombinant human microplasminogen and pilot study in inducing posterior vitreous detachment[J]. CURRENT EYE RESEARCH,2005,30(10):881-889.
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