کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10843541 | 1069270 | 2005 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Production and characterization of a bacterial single-chain antibody fragment specific to B-cell-activating factor of the TNF family
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Production and characterization of a bacterial single-chain antibody fragment specific to B-cell-activating factor of the TNF family Production and characterization of a bacterial single-chain antibody fragment specific to B-cell-activating factor of the TNF family](/preview/png/10843541.png)
چکیده انگلیسی
An active form of a single-chain antibody fragment (scFv) from the murine monoclonal antibody ABL-1, which is specific for B-cell-activating factor of the TNF family, was produced in Escherichia coli. The complementary DNAs encoding the variable regions of the heavy chain (VH) and light chain (VL) were connected by a (Gly4Ser)3 linker, using an assembly polymerase chain reaction. The construct VH-linker-VL was placed under the control of highly efficient T7 promoter system. The cloned scFv was expressed in E. coli BL21(DE3) as inclusion bodies. After extraction from the E. coli cells, the inclusion bodies were solubilized and denatured in the presence of 8 M urea. The expressed scFv fusion proteins were purified by Ni2+-IDA His-bind resin and finally renatured by dialysis. The purity and activity of the purified scFv were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blotting, and enzyme-linked immunosorbent assay. The result revealed that the ABL-1 scFv retains the specific binding activity to BAFF with an affinity constant of 0.9 Ã 10â8 mol Lâ1.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 43, Issue 2, October 2005, Pages 157-164
Journal: Protein Expression and Purification - Volume 43, Issue 2, October 2005, Pages 157-164
نویسندگان
Peng Cao, Xiang Ming Tang, Zheng Bing Guan, Zhen Yu Diao, Shuang Quan Zhang,