TY - JOUR
T1 - Chromatographic biopanning for the selection of peptides with high specificity to Pb2+ from phage displayed peptide library
AU - Nian, Rui
AU - Kim, Duck Sang
AU - Nguyen, Thuong
AU - Tan, Lihan
AU - Kim, Chan Wha
AU - Yoo, Ik Keun
AU - Choe, Woo Seok
N1 - Funding Information:
This work was supported by Korea Institute of Environmental Science and Technology (KIEST, Grant no. 2008-10001-0029-0 ).
PY - 2010/9
Y1 - 2010/9
N2 - 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.
AB - 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.
KW - Chromatographic biopanning
KW - FPLC
KW - Monolithic column
KW - Pb
KW - Peptide
KW - Phage display
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U2 - 10.1016/j.chroma.2010.07.048
DO - 10.1016/j.chroma.2010.07.048
M3 - Article
C2 - 20709321
AN - SCOPUS:77956180001
SN - 0021-9673
VL - 1217
SP - 5940
EP - 5949
JO - Journal of Chromatography A
JF - Journal of Chromatography A
IS - 38
ER -