Peptide Microarray with Ligands at High Density Based on Symmetrical Carrier Landscape Phage for Detection of Cellulase

CORC > 青岛生物能源与过程研究所 > 中国科学院青岛生物能源与过程研究所 > 生物传感技术团队

	Peptide Microarray with Ligands at High Density Based on Symmetrical Carrier Landscape Phage for Detection of Cellulase
	Qi, Huan 1,2,3; Wang, Fei 1,2,3; Petrenko, Valery A.4; Liu, Aihua 1,2,3
刊名	ANALYTICAL CHEMISTRY
	2014-06-17
卷号	86 期号:12 页码:5844-5850
英文摘要	Peptide microarrays evolved recently as a routine analytical implementation in various research areas due to their unique characteristics. However, the immobilization of peptides with high density in each spot during the fabricating process remains a problem, which will affect the performance of the resultant microarray greatly. To respond to this challenge, a novel peptide immobilization method using symmetrical phage carrier was developed in this work. The cellulytic enzyme endoglucanase I (EG I) was used as a model for selection of its specific peptide ligands from the f8/8 landscape library. Three phage monoclones were selected and identified by the specificity array, of which one phage monoclone displaying the fusion peptide EGSDPRNIV (phage EGSDPRMV) could bind EG I specifically with highest affinity. Subsequently, the phage EGSDPRMV was used directly to construct peptide microarray. For comparison, major coat protein pVIII fused EG I specific peptide EGSDPRMV (pVIII-fused EGSDPRMV) which was isolated from phage EGSDPRMV was also immobilized by traditional method to fabricate peptide microarray. The fluorescent signal of the phage EGSDPRMV-mediated peptide microarray was more reproducible and about four times higher than the value for pVIII-fused EGSDPRMV-based microarray, suggesting the high efficiency of the proposed phage EGSDPRMV-mediated peptide immobilization method. Further, the phage EGSDPRMV based microarray not only simplified the procedure of microarray construction but also exhibited significantly enhanced sensitivity due to the symmetrical carrier landscape phage, which dramatically increased the density and sterical regularity of immobilized peptides in each spot. Thus, the proposed strategy has the advantages that the immobilizing peptide ligands were not disturbed by their composition and the immobilized peptides were highly regular with free amino-terminal.
WOS标题词	Science & Technology ; Physical Sciences
类目[WOS]	Chemistry, Analytical
研究领域[WOS]	Chemistry
关键词[WOS]	BREAST-CANCER CELLS ; PROTEIN IMMOBILIZATION ; DISPLAY ; PROBES ; SELECTION ; LIBRARY ; CHIPS ; POLYPEPTIDES ; SPECIFICITY ; RECOGNITION
收录类别	SCI
语种	英语
WOS记录号	WOS:000337643500038
公开日期	2015-12-24
内容类型	期刊论文
源URL	[http://ir.qibebt.ac.cn/handle/337004/6216]
专题	青岛生物能源与过程研究所_生物传感技术团队
作者单位	1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Lab Biosensing, Qingdao 266101, Peoples R China 2.Chinese Acad Sci, Key Lab Bioenergy, Qingdao 266101, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Auburn Univ, Dept Pathobiol, Auburn, AL 36849 USA
推荐引用方式 GB/T 7714	Qi, Huan,Wang, Fei,Petrenko, Valery A.,et al. Peptide Microarray with Ligands at High Density Based on Symmetrical Carrier Landscape Phage for Detection of Cellulase[J]. ANALYTICAL CHEMISTRY,2014,86(12):5844-5850.
APA	Qi, Huan,Wang, Fei,Petrenko, Valery A.,&Liu, Aihua.(2014).Peptide Microarray with Ligands at High Density Based on Symmetrical Carrier Landscape Phage for Detection of Cellulase.ANALYTICAL CHEMISTRY,86(12),5844-5850.
MLA	Qi, Huan,et al."Peptide Microarray with Ligands at High Density Based on Symmetrical Carrier Landscape Phage for Detection of Cellulase".ANALYTICAL CHEMISTRY 86.12(2014):5844-5850.