Analysis of low molecular weight plasma proteins using ultrafiltration and large gel two-dimensional electrophoresis

Woon Won Jung, Sohee Phark, Sangnam Oh, Jin Young Khim, Juneyoung Lee, Myung Hee Nam, Jong Bok Seo, So Young Park, Eunkyung Jo, Seonyoung Choi, Zhi Zheng, Ji Young Lee, Min Lee, Eunil Lee, Donggeun Sul

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Abstract

In this study, various solvent systems were applied to obtain a high and consistent recovery rate of low molecular weight plasma proteins (LMPP) from human plasma. A buffer system containing 7 M urea, 2 M thiourea, 25 mM NH 4HCO3 1 20% ACN (pH 8.2) produced the highest recovery rate of LMPP. To validate the recovery of cut off membrane (COM) obtained using the urea buffer system, 27 different 30 kDa COMs were used to prepare the LMPP sample which were then subjected to 1-D SDS-PAGE. Statistical analysis showed that the buffer system with COM produced a consistent the recovery of LMPP. In addition, 2-DE analysis was also conducted to determine the relative intensity of each protein spot. When molecular weight ranges over 30 kDa and under 30 kDa were evaluated, 953 and 587 protein spots were observed in the gels, respectively, resulting in a total of 1540 protein spots being resolved. Identification of the major proteins were then performed using a nano-LC/MS system comprised of an HPLC system and an ESI-quadrupole IT MS equipped with a nano-ESI source.

Original languageEnglish
Pages (from-to)1827-1840
Number of pages14
JournalProteomics
Volume9
Issue number7
DOIs
Publication statusPublished - 2009 Apr 1

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Keywords

  • Acetonitrile
  • Cut off membranes
  • Immobilized pH gradients
  • Low molecular weight plasma proteins
  • Major high molecular weight plasma proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Jung, W. W., Phark, S., Oh, S., Khim, J. Y., Lee, J., Nam, M. H., Seo, J. B., Park, S. Y., Jo, E., Choi, S., Zheng, Z., Lee, J. Y., Lee, M., Lee, E., & Sul, D. (2009). Analysis of low molecular weight plasma proteins using ultrafiltration and large gel two-dimensional electrophoresis. Proteomics, 9(7), 1827-1840. https://doi.org/10.1002/pmic.200800470