Effects of signal sequence on phage-displayed disulfide-stabilized single chain antibody variable fragment (sc-dsFv) libraries

Yu Ching Lee, Ing Chien Chen, Chung Ming Yu, Yi Jen Huang, Hung Ju Hsu, An Suei Yang

研究成果: 雜誌貢獻文章同行評審

摘要

Phage-displayed single chain variable fragment (scFv) libraries are powerful tools in antibody engineering. Disulfide-stabilized scFv (sc-dsFv) with an interface disulfide bond is structure-wise more stable than the corresponding scFv. A set of recently discovered signal sequences replacing the wild type (pelB) signal peptidase cleavage site in the c-region has been shown to be effective in rescuing the expression of sc-dsFv libraries on the phage surface. However, the effects of the other regions of the signal sequence on the expression of the sc-dsFv libraries and on the formation of the interface disulfide bond in the phage-displayed sc-dsFv have not been clear. In this work, selected novel signal sequence variants in the h-region were shown to be equally effective in promoting sc-dsFv library expression on the phage surface; the expression level and complexity of the sc-dsFv libraries were comparable to the corresponding scFv libraries produced with the wild-type (pelB) signal sequence. The interface disulfide bond in the phage-displayed sc-dsFv was proven to form to a large extent in the library variant ensemble generated with signal sequence variants in both the h-region and the c-region. The sc-dsFv engineering platform established in this work can be applied to many of the known scFv molecules which are in need of a more stable version for the applications under harsh conditions or for longer shelf-life.
原文英語
頁(從 - 到)348-353
頁數6
期刊Biochemical and Biophysical Research Communications
411
發行號2
DOIs
出版狀態已發佈 - 七月 29 2011
對外發佈

ASJC Scopus subject areas

  • 生物化學
  • 生物物理學
  • 細胞生物學
  • 分子生物學

指紋

深入研究「Effects of signal sequence on phage-displayed disulfide-stabilized single chain antibody variable fragment (sc-dsFv) libraries」主題。共同形成了獨特的指紋。

引用此