Chinese Journal of Chromatography ›› 2014, Vol. 32 ›› Issue (6): 600-603.DOI: 10.3724/SP.J.1123.2014.03031

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Preliminary study of two steps aqueous two-phase extraction combined with high performance liquid chromatography for saliva protein selective separation

ZHAO Xinying1,2, QU Feng1, QIN Hao3, LUO Aiqin1   

  1. 1. School of Life Science, Beijing Institute of Technology, Beijing 100081, China;
    2. Beijing Center for Physical and Chemical Analysis, Beijing 100089, China;
    3. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
  • Received:2014-03-21 Revised:2014-03-28 Online:2014-06-08 Published:2014-05-22
  • Supported by:

    National Basic Research Program of China (973 Program) (2012CB910603); National Natural Science Foundation of China (21175011, 21375008).

Abstract:

Saliva protein separation by one-step aqueous two-phase extraction (ATPE) in the system of 15% (mass ratio if not specified) PEG-4000/8% NaH2PO4 was presented. The proteins extracted from the top and bottom phases were separated and characterized by high performance liquid chromatography (HPLC) respectively. Under the optimized gradient elution procedure, more saliva protein peaks in the two phases were shown. Saliva protein peaks in 50 min gradient were divided into ten specific groups which were classified into six sections based on the protein distribution in the top and bottom phases. Then, the top or bottom phase was performed the second step ATPE. Possible reasons for protein partition and retention difference are discussed briefly. Results show that some proteins can be separated completely in one-step ATPE, while some can be separated in the following two-step ATPE. The combination of two steps ATPE and HPLC provides a new way to achieve selective separation.

Key words: aqueous two-phase extraction (ATPE), high performance liquid chromatography (HPLC), saliva protein, two-step extraction

CLC Number: