Chromatographic biopanning for the selection of peptides with high specificity to Pb2+ from phage displayed peptide library

Rui Nian, Duck Sang Kim, Thuong Nguyen, Lihan Tan, Chan Wha Kim, Ik Keun Yoo, Woo Seok Choe

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Toxic heavy metal pollution is a global problem occurring in air, soil as well as water. There is a need for a more cost effective, renewable remediation technique, but most importantly, for a recovery method that is selective for one specific metal of concern. Phage display technology has been used as a powerful tool in the discovery of peptides capable of exhibiting specific affinity to various metals or metal ions. However, traditional phage display is mainly conducted in batch mode, resulting in only one equilibrium state hence low-efficiency selection. It is also unable to monitor the selection process in real time mode. In this study, phage display technique was incorporated with chromatography procedure with the use of a monolithic column, facilitating multiple phage-binding equilibrium states and online monitoring of the selection process in search of affinity peptides to Pb2+. In total, 17 candidate peptides were found and their specificity toward Pb2+ was further investigated with bead-based enzyme immunoassay (EIA). A highly specific Pb2+ binding peptide ThrAsnThrLeuSerAsnAsn (TNTLSNN) was obtained. Based on our knowledge, this is the first report on a new chromatographic biopanning method coupled with monolithic column for the selection of metal ion specific binding peptides. It is expected that this monolith-based chromatographic biopanning will provide a promising approach for a high throughput screening of affinity peptides cognitive of a wide range of target species.

Original languageEnglish
Pages (from-to)5940-5949
Number of pages10
JournalJournal of Chromatography A
Volume1217
Issue number38
DOIs
Publication statusPublished - 2010 Sep

Keywords

  • Chromatographic biopanning
  • FPLC
  • Monolithic column
  • Pb
  • Peptide
  • Phage display

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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