• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to project page

1994 Fiscal Year Final Research Report Summary

Separation of Biopolymers by Countercurrent Chromatography

Research Project

Project/Area Number 05671792
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Physical pharmacy
Research InstitutionTokyo University of Pharmacy and Life Science

Principal Investigator

SHIBUSAWA Yoichi  Tokyo University of Pharmacy and Life Science, School of Pharmacy, Lecturer, 薬学部, 講師 (10102708)

Project Period (FY) 1993 – 1994
KeywordsCountercurrent Chromatography / Human Serum Lipoproteins / Aqueous Polymer Two Phase Systems / DNA Binding Proteins
Research Abstract

Three main classes lipoproteins from human serum, one step purification of single stranded binding protein (SSB) and H-NA from E.coli were studied by countercurrent chromatography.
High- and low-density lipoproteins (HDLs, LDLs) were fractionated by countercurrent chromatography using the cross-axis coil planet centrifuge using two-phase aqueous polymer systems. The best separations were achieved with a polymer phase system composed of 16% polyethylene glycol 1000 and 12.5% dibasic potassium phosphate at pH 9.4.HDL-LDL fractions were collected from 4 ml of human serum within 3h. Very-low-lipoproteins (VLDLs) were collected together with serum proteins. Each fraction was concentrated and subjected to hydroxyapatite chromatography to obtain three lipoprotein fractions (HDLs, LDLs and VLDLs) all free of serum proteins, such as albumin and globulin. Each lipoprotein was confirmed by agarose gel electrophoresis.
SSB purification from crude extracts of E.coli was performed by cross-axis coil planet centrifuge using aqueous polymer-two phase system composed of 16% PEG 1000-17% ammonium sulfate. Countercurrent chromatogram gives two main peaks, one contains most of proteins produced by E.coli, and the other one (fraction 51) contains SSB.SSB was confirmed by SDS-PAGE.

  • Research Products

    (4 results)

All Other

All Publications (4 results)

  • [Publications] Y.Shibusawa et al.: "Countercurrent Chromatographic Isolation of High- and Low Density Lipoprotein Fractions from Human Serum" Countercurrent Chromatography (ACS Mono-graphs). in press.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Shibusawa et al.: "Complementary Use of Counter-Current Chromatography and Hydroxyapatite Chromatography for the Separation of Three Main Classes of Lipoproteins from Human Serum" Journal of Chromatography, Biomedical Appl.664. 295-301 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Shibusawa, T.Chiba, U.Matsumoto and Y.Ito: "Countercurrent Chromatographic Isolation of High- and Low-Density Lipoprotein Fractions from Human Serum" Countercurrent Chromatography (ACS Monographs). (in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Shibusawa, M.Mugiyama, U.Matsumoto and Y.Ito: "Complementary Use of Counter-Current Chromatography and Hydroxyapatite Chromatography for the Separation of Three Main Classes of Lipoproteins from Human Serum" Journal of Chromatography Biomedical Applications. 664. 295-301 (1995)

    • Description
      「研究成果報告書概要(欧文)」より

URL: 

Published: 1996-04-15  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi