检索结果-南通市图书馆

Construction and expression of a bicistronic vector containing human bone morphogenetic protein 2 and vascular endothelial growth factor-165 genes in vitro

引用

Chinese Medical Journal 2009年第4期122卷 471-473页

作者： TIAN Xiao-bin SUN Li ZHANG Yu-kun GAO Yong FU De-hao YANG Shu-hua Department of Orthopedics People's Hospital of GuizhouProvince Guiyang Guizhou 550002 China Department of Orthopedics Union Hospital Tongji MedicalCollege of Huazhong Science and Technology UniversityWuhan Hubei 430022 China

It has been well documented that bone morphogenetic .proteins （BMPs）, a group of proteins belonging to the transforming growth factor-β （TGFβ） superfamily, can induce bone formation, both in vivo and in vitro. Bone morphogenetic protein-2 （BMP2） is a potent osteoinductive factor and is being evaluated as a bone growth inducer for orthopedic applications.1 Vascular endothelial growth factor （VEGF）, the best-characterized angiogenic factor,

关键词： bone morphogenetic protein 2 vascular endothelial growth factor-165 co-expression bicistronic vector

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Functionality of a bicistronic construction containing HEXA and HEXB genes encoding β-hexosaminidase A for cell-mediated therapy of GM2 gangliosidoses

引用

Neural Regeneration Research 2022年第1期17卷 122-129页

作者： Alisa A.Shaimardanova Daria S.Chulpanova Valeriya V.Solovyeva Aleksandr M.Aimaletdinov Albert A.Rizvanov Institute of Fundamental Medicine and Biology Kazan Federal UniversityKazanRussia

Tay-Sachs disease and Sandhoff disease are severe hereditary neurodegenerative disorders caused by a deficiency ofβ-hexosaminidase A(HexA)enzyme,which results in the accumulation of GM2 gangliosides in the nervous system cells.In this work,we analyzed the efficacy and safety of cell-mediated gene therapy for Sandhoff disease and Sandhoff disease using a bicistronic lentiviral vector encoding cDNA of HexAα-andβ-subunit genes separated by the nucleotide sequence of a P2A peptide(HEXA-HEXB).The functionality of the bicistronic construct containing the HEXA-HEXB genetic cassette was analyzed in a culture of HEK293T cells and human umbilical cord blood mononuclear cells(hUCBMCs).Our results showed that the enzymatic activity of HexA in the conditioned medium harvested from genetically modified HEK293T-HEXA-HEXB and hUCBMCs-HEXA-HEXB was increased by 23 and 8 times,respectively,compared with the conditioned medium of native cells.Western blot analysis showed that hUCBMCs-HEXA-HEXB secreted both completely separated HEXA and HEXB proteins,and an uncleaved protein containing HEXA+HEXB linked by the P2A peptide.Intravenous injection of genetically modified hUCBMCs-HEXA-HEXB to laboratory Wistar rats was carried out,and the HexA enzymatic activity in the blood plasma of experimental animals,as well as the number of live cells of immune system organs(spleen,thymus,bone marrow,lymph nodes)were determined.A significant increase in the enzymatic activity of HexA in the blood plasma of laboratory rats on days 6 and 9(by 2.5 and 3 times,respectively)after the administration of hUCBMCsHEXA-HEXB was shown.At the same time,the number of live cells in the studied organs remained unchanged.Thus,the functionality of the bicistronic genetic construct encoding cDNA of the HEXA and HEXB genes separated by the nucleotide sequence of the P2A peptide was shown in vitro and in vivo.We hypothesize that due to the natural ability of hUCBMCs to overcome biological barriers,such a strategy can restore the activity of the missing enzyme in the ce

关键词： bicistronic vector cell-mediated gene therapy GM2 gangliosidosis P2A peptide Sandhoff disease Tay-Sachs disease umbilical cord blood mononuclear cells β-hexosaminidase

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Intron Enhances Humanized Antibody anti-PD-1 Expression

Intron Enhances Humanized Antibody anti-PD-1 Expression

引用

第十届全国免疫学学术大会

作者： Jie Liu 中国科学院深圳先进技术研究院

Background:Monoclonal antibody(m Ab)is used more and more widely in biopharmaceuticals at cancer and other difficult miscellaneous diseases.Recently,the clinical development therapy based on the PD-1 has resulted in a major breakthrough in the field of oncology.However how to produce high quality antibody for clinical trial is still a major problem.Purpose:In this study we designed a set of bicistronic vector for eukaryotic expression in 293F cell line that produced both selected marker and recombinant c DNA by controlled in two different promoters.Hoping to use this expression system,we can humanized the mouse antibody and can improve the antibody expression to satisfy the application for clinical trial.Methods:Firstly,we used the pc DNA3.1(+)as the backbone to construct the bicistronic vector,by replacing the neomycin to the GS sequences and successively inserting the IRES,the CH of heavy chain and intron.After insert the variable region of the murine antibody,we used the PEI transfection method to expression the anti-PD-1 antibody in 293F cell line.And the supernatant was used to testify the antibody expression by SDS-PAGE and Western Blot.Then we used the protein A agarose to purify the antibody,and detected the binding activity and specificity of the antibody by Elisa.Results:In this expression system,we found thatβ-globin intron followed with promoter can dramatically enhance the expression of humanized anti-PD1 antibody.As compared to the non-intron control,intron significantly increase the antibody yield b at least 5 folds without affecting the special affinity to the antigen PD-1.Conclusion:The intron-mediated bicistronic vector can remarkable improve the antibody expression,which provides an attractive tool for fast generation of m Ab in 293F cell lines with high productivity.

关键词： Anti-PD-1 antibody bicistronic vector Intron Elisa

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：