Chinese Journal of Chromatography ›› 2016, Vol. 34 ›› Issue (12): 1264-1270.DOI: 10.3724/SP.J.1123.2016.09033

• Articles • Previous Articles    

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

TANG Jun1,2,5, GUO Kaizhu2, CHENG Wendong2, SONG Peipei3, FENG Shun3, HU Chaofeng5, XU Ruilian1, TIAN Ruijun2,4   

  1. 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:2016-09-15 Online:2016-12-08 Published:2013-04-24
  • Supported by:

    National Natural Science Foundation of China (No. 21575057); Shenzhen Innovation of Science and Technology Commission (Nos. JCYJ20150901153557178, JSGG20160301103415523).

Abstract:

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.

Key words: Fe3O4/EDTA (ethylenediaminetetraacetic acid), magnetic beads, proteomics, sample preparation

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