An Improved Method for Removal of Antithrombin III from Post-heparin Plasma Lipase Fractions with a High Recovery Rate

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概要

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• Affinity chromatography on heparin-Sepharose has been widely used for the purification of postheparin plasma triglyceride lipases, but this procedure alone yields lipase fractions with a high content of antithrombin III (AT), which also binds to heparin and coelutes with the lipases. A rapid procedure in which this major contaminant is reduced by the use of affinity chromatography on heparin-Sepharose with low affinity for AT and further removal by anti-AT IgG Affi-Gel 10 column chromatography is reported and the results are compared with these of the conventional heparin-Sepharose and Concanavalin A-Sepharose column chromatography two step method. While the AT contamination of hepatic lipase eluate from conventional heparin-Sepharose column chromatography was 92 mg/dl in 100 mg/dl protein, heparin-Sepharose with low affinity for AT yielded samples with 55 mg/dl in 100 mg/dl protein. A second passage on concanavalin A-Sepharose of the sample from conventional heparin-Sepharose reduced the AT content from 92 mg/dl to 42 mg/dl in 100 mg/dl protein, but passage of the sample from heparin-Sepharose with low affinity for AT through an anti-AT IgG Affi-Gel 10 column removed almost all of the AT contamination from the lipase sample. This hepatic triglyceride lipase (HTGL) sample had a specific activity 1,963-fold higher than that of postheparin plasma with a yield of 17% of the starting material, showing an excellent recovery rate compared with similar methods in use.

• 東海大学の論文

著者

• GOTO Yuichiro

  Department of Internal Medicine Keio University

• Homma Yasuhiko

  Department Of Internal Medicine Tokai University School Of Medicine

• Homma Yasuhiko

  Department Of Clinical Health Science Tokai University School Of Medicine

• Goto Yuichiro

  Tokai University School Of Medicine Department Of Internal Medicine

• Goto Yuichiro

  Dept Of Intern. Med Keio Univ. School Of Med.

• Goto Yuichiro

  東海大学 循環器科

• Tamachi Hiromitsu

  Department of Internal Medicine, Tokai University School of Medicine

• Honma Yasuhiko

  School Of Med. Keio Univ

• Goto Yuichiro

  Department Of 1st Internal Medicine Tokai University School Of Medicine

• Honma Yasuhiko

  Department Of Internal Medicine Tokai University School Of Medicine

• MORIGUCHI Emilio

  Department of Internal Medicine, Tokai University School of Medicine

• Gotoh Yuichiro

  First Department Of Internal Medicine School Of Medicine Tokai University

• Tada Hiroki

  Department Of Internal Medicine Tokai University

• Tamachi H

  Tokai Univ. School Of Medicine Jpn

• Gotoh Yuichiro

  Department Of Internal Medicine Tokai University School Of Medicine

• Homma Y

  Department Of Internal Medicine Tokai University School Of Medicine

• Goto Y

  Kobe Steel. Ltd. Kobe‐shi Jpn

• Tamachi Hiromitsu

  Department Of Internal Medicine Tokai University School Of Medicine

• Goto Yuichiro

  First Department Of Internal Medicine Tokai University School Of Medicine Japan

• Tomoda Haruo

  Department Of Cardiology

• Moriguchi E

  Tokai Univ. School Of Medicine Jpn

• Tomoda Haruo

  Department Of Internal Medicine Tokai University School Of Medicine

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