Citation: TANG Jun, GUO Kaizhu, CHENG Wendong, SONG Peipei, FENG Shun, HU Chaofeng, XU Ruilian, TIAN Ruijun. Fe₃O₄/ethylenediaminetetraacetic acid magnetic beads-based integrated method for proteomic analysis[J]. Chinese Journal of Chromatography, ;2016, 34(12): 1264-1270. doi: 10.3724/SP.J.1123.2016.09033 shu

Fe₃O₄/ethylenediaminetetraacetic acid magnetic beads-based integrated method for proteomic analysis

TANG Jun ^1,2,5, ,
GUO Kaizhu ² ,
CHENG Wendong ² ,
SONG Peipei ³ ,
FENG Shun ³ ,
HU Chaofeng ⁵ ,
XU Ruilian ¹ ,
TIAN Ruijun ^2,4

1.
Institute of Oncology, Shenzhen People's Hospital, Shenzhen 518020, China;
2.
Department of Chemistry, South University of Science and Technology of China, Shenzhen 518055, China;
3.
School of Life Science and Engineering, Southwest Jiaotong University, Chengdu 610031, China;
4.
Shenzhen Key Laboratory of Cell Microenvironment, Shenzhen 518055, China;
5.
Department of Pathophysiology, School of Basic Medicine, Jinan University, Guangzhou 510632, China

Received Date: 15 September 2016

Fund Project: National Natural Science Foundation of China (No. 21575057)

A Fe₃O₄/EDTA (ethylenediaminetetraacetic acid) magnetic beads-based integrated method for proteomic analysis was developed. Fe₃O₄ magnetic beads loaded with EDTA were prepared firstly via co-precipitation method. One hundred μ g Fe₃O₄/EDTA magnetic beads could adsorb 12.4 μ g BSA (bovine serum albumin) in the optimized solution (95% acetonitrile-1% trifluoracetic acid, v/v). The binding capacity was about 10 times higher than that of the commercialized magnetic beads. The digestion time of Fe₃O₄/EDTA magnetic beads as the proteomic reactor was also optimized using BSA as the standard protein. The results indicated that the peptide coverage and unique peptides obtained from Fe₃O₄/EDTA magnetic beads with digestion time of 1, 8 and 16 h were comparable. We then applied the Fe₃O₄/EDTA magnetic beads for the serum proteome analysis. A total of 218 proteins were identified successfully, including 41 biomarkers approved by Food and Drug Administration (FDA). Protein preconcentration, reduction, alkylation, digestion, peptide desalting, and elution can be achieved in an integrated manner by the Fe₃O₄/EDTA magnetic beads-based proteomics sample preparation, which reduced the sample loss during sample transfering and processing. The developed method is fast, sensitive and easy to operate, which has prospective application for the clinic proteomics research.
- Fe₃O₄/EDTA (ethylenediaminetetraacetic acid),
- magnetic beads,
- proteomics,
- sample preparation
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Fe3O4/ethylenediaminetetraacetic acid magnetic beads-based integrated method for proteomic analysis

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通讯作者: 陈斌, bchen63@163.com

Fe₃O₄/ethylenediaminetetraacetic acid magnetic beads-based integrated method for proteomic analysis