A human single-chain variable fragment targeting to Vibrio vulnificus RtxA toxin

Fang Yu Hsu, Li Fang Chou, Lien I. Hor, Hwan You Chang*

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

8 Scopus citations

Abstract

Vibrio vulnificus secretes a multifunctional cytotoxin RtxA (VvRtxA), which plays a major role in the bacterial pathogenesis. The lack of an efficient VvRtxA detection tool has hampered the progress of V. vulnificus pathogenesis research. This study aims to isolate VvRtxA specific single-chain variable fragments (scFv) to serve as a detection agent. The VvRtxA C-terminal Gly-Asp (GD) repeat-containing region, which has been implicated for calcium binding and target cell recognition, was chosen as an antigen to screen a scFv phage display library. A scFv clone that reacted positively to VvRtxA was successfully obtained. Using the isolated scFv, a cell-based enzyme-linked immunosorbent assay was established for detecting cell-associated VvRtxA toxin in V. vulnificus infected HEp-2 cells. The result is consistent with previous observations that secretion of VvRtxA toxin is time-dependent on bacteria contacting with host cells. Utilization of scFv for VvRtxA toxin detection provides an applicable strategy devoid of conventional immunization.

Original languageEnglish
Pages (from-to)94-100
Number of pages7
JournalJournal of Microbiological Methods
Volume84
Issue number1
DOIs
StatePublished - 01 2011
Externally publishedYes

Keywords

  • Gly-Asp rich region
  • RtxA
  • Single-chain variable fragment
  • Vibrio vulnificus

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