دورية أكاديمية
Production of recombinant human microplasminogen and pilot study in inducing posterior vitreous detachment
| العنوان: | Production of recombinant human microplasminogen and pilot study in inducing posterior vitreous detachment |
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| المؤلفون: | Qu, JF, Li, XX, Huang, BR |
| المساهمون: | Li, XX (reprint author), Peking Univ, Peoples Hosp, Beijing 100871, Peoples R China., Peking Univ, Peoples Hosp, Beijing 100871, Peoples R China., Peking Union Med Coll, Beijing, Peoples R China. |
| المصدر: | PubMed ; SCI |
| بيانات النشر: | current eye research |
| سنة النشر: | 2005 |
| المجموعة: | Peking University Institutional Repository (PKU IR) / 北京大学机构知识库 |
| مصطلحات موضوعية: | Pichia pastoris, posterior vitreous detachment, recombinant human microplasminogen, PLASMIN-ASSISTED VITRECTOMY, DIFFERENT MOLECULAR FORMS, MACULAR HOLES, VITREORETINAL INTERFACE, TRACTION SYNDROME, ACTIVE-SITE, STREPTOKINASE, ACTIVATION, EXPRESSION, UROKINASE |
| الوصف: | 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. ; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000233090300009&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701Test ; Ophthalmology ; SCI(E) ; PubMed ; 2 ; ARTICLE ; 10 ; 881-889 ; 30 |
| نوع الوثيقة: | journal/newspaper |
| اللغة: | English |
| ردمك: | 978-0-00-233090-9 0-00-233090-3 |
| تدمد: | 0271-3683 |
| العلاقة: | CURRENT EYE RESEARCH.2005,30,(10),881-889.; 675281; http://hdl.handle.net/20.500.11897/199049Test; WOS:000233090300009 |
| DOI: | 10.1080/02713680591006282 |
| الإتاحة: | https://doi.org/20.500.11897/199049Test https://doi.org/10.1080/02713680591006282Test https://hdl.handle.net/20.500.11897/199049Test |
| رقم الانضمام: | edsbas.D1148103 |
| قاعدة البيانات: | BASE |
| ردمك: | 9780002330909 0002330903 |
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| تدمد: | 02713683 |
| DOI: | 10.1080/02713680591006282 |