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

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
卷411
發行號2
DOIs
出版狀態已發佈 - 七月 29 2011
對外發佈Yes

指紋

Single-Chain Antibodies
Bacteriophages
Protein Sorting Signals
Disulfides
Libraries
Molecules
Antibodies

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

引用此文

Effects of signal sequence on phage-displayed disulfide-stabilized single chain antibody variable fragment (sc-dsFv) libraries. / Lee, Yu Ching; Chen, Ing Chien; Yu, Chung Ming; Huang, Yi Jen; Hsu, Hung Ju; Yang, An Suei.

於: Biochemical and Biophysical Research Communications, 卷 411, 編號 2, 29.07.2011, p. 348-353.

研究成果: 雜誌貢獻 › 文章

Lee, YC, Chen, IC, Yu, CM, Huang, YJ, Hsu, HJ & Yang, AS 2011, 'Effects of signal sequence on phage-displayed disulfide-stabilized single chain antibody variable fragment (sc-dsFv) libraries', Biochemical and Biophysical Research Communications, 卷 411, 編號 2, 頁 348-353. https://doi.org/10.1016/j.bbrc.2011.06.146
Lee YC, Chen IC, Yu CM, Huang YJ, Hsu HJ, Yang AS. Effects of signal sequence on phage-displayed disulfide-stabilized single chain antibody variable fragment (sc-dsFv) libraries. Biochemical and Biophysical Research Communications. 2011 7月 29;411(2):348-353. https://doi.org/10.1016/j.bbrc.2011.06.146
Lee, Yu Ching ; Chen, Ing Chien ; Yu, Chung Ming ; Huang, Yi Jen ; Hsu, Hung Ju ; Yang, An Suei. / Effects of signal sequence on phage-displayed disulfide-stabilized single chain antibody variable fragment (sc-dsFv) libraries. 於: Biochemical and Biophysical Research Communications. 2011 ; 卷 411, 編號 2. 頁 348-353.
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