Chemical Research in Chinese Universities ›› 2010, Vol. 26 ›› Issue (1): 17-22.

• Articles • Previous Articles     Next Articles

Rapid Purification and Identification of Osteopontin from Human Milk by HPLC Coupled to FT-ICR-MS

SHI Lei1*, YANG Tie-zheng2 and ZUBAREV A. Roman3   

  1. 1. College of Public Health, Jilin University, Changchun 130021, P. R. China;
    2. Affiliated Hospital of Changchun University of Chinese Medicine, Changchun 130021, P. R. China;
    3. Laboratory for Biological and Medical Mass Spectrometry, BMC, Uppsala University, Box 583, 75123 Uppsala, Sweden
  • Received:2009-05-12 Revised:2009-08-03 Online:2010-01-04 Published:2010-03-29
  • Contact: SHI Lei. E-mail: shilei@jlu.edu.cn
  • Supported by:

    Supported by the Project from Scientific Bureau of Jilin Province, China(No.200705391).

Abstract:

Milk as a key element for infant nutrition represents the only source of feeding for newborns and infants, breast-feeding milk normally contains several bioactive proteins or peptides useful for the development of the immune system that protects infants from diseases. Osteopontin(OPN) plays a distinct role during the processes of lactation. Some studies on OPN isolated and purified from the human milk via HPLC on SCX and C4 columns adopted biological mass spectrometry. After digesting the purified OPN sample with trypsin, the typical correlative polypeptide fragments GDSVVYGLR and QNLLAPQTLPSK are identified by FT-ICR-MS. The rapid identification of OPN is assumed to be reasonably well behaving in a standard bottom-up experiment. It shows that nano-spray HPLC combined with FT-ICR-MS and Mascot search can be used as a high efficient method for the identification of purified OPN and its polypeptide fragments. Further study will provide more academic theories of their different modifications and the bioactivity as well.

Key words: Osteopontin(OPN); FT-ICR-MS; Purification; Identification