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Talanta. 2015 Mar;134:705-711. doi: 10.1016/j.talanta.2014.12.012. Epub 2014 Dec 18.

Label-free detection microarray for novel peptide ligands screening base on MS-SPRi combination.

Author information

1
CAS Key Laboratory for Biomedical Effects of Nanomaterials & Nanosafety, National Center for Nanoscience and Technology of China, Beijing 100190, China. Electronic address: wangwz@nanoctr.cn.
2
CAS Key Laboratory for Biomedical Effects of Nanomaterials & Nanosafety, National Center for Nanoscience and Technology of China, Beijing 100190, China.
3
CAS Key Laboratory for Biomedical Effects of Nanomaterials & Nanosafety, National Center for Nanoscience and Technology of China, Beijing 100190, China; Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China; Institute for Systems Biology, 401 Terry Avenue N, Seattle, WA 98109, USA. Electronic address: huzy@nanoct.cn.

Abstract

Peptides ligands with high affinity and high specificity towards specific targets is catching a good deal of interests in biomedical field. Traditional peptide screening procedure involves selection, sequencing and characterization and each step is time-consuming and labor-intensive. The combination between different analytical methods could provide an integrated plan for efficient peptide screening. We report herein a label-free detection microarray system to facilitate the whole one-bead-one-compound (OBOC) peptide screening process. A microwell array chip with two identical units can trap the candidate peptide beads in one-well-one-bead manner. Peptides on beads were photo-released in situ in the well and partly transferred to two identical chips for Surface Plasmon Resonance imaging (SPRi), and peptide left in the bi-unit microwell array chip was remain for in situ single bead sequencing by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Using the bi-unit imprinted chip system, affinity peptides towards AD protein were efficiently screened out both qualitatively and quantitatively from 10(4) candidates. The method provides a universal solution for high efficiency and high throughput ligands screening.

KEYWORDS:

Bi-unit; Label-free detection; MALDI-TOF-MS; Microarray; SPRi

PMID:
25618725
DOI:
10.1016/j.talanta.2014.12.012
[Indexed for MEDLINE]

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