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Development of analytical methods by capillary electophoresis for human blood isoenzymes.

Research Project

Project/Area Number 07457617
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory medicine
Research InstitutionKumamoto University

Principal Investigator

UJI Yoshinori  Ph.D., Kumamoto University, School of Medicine, Assistant Professor, 医学部, 助手 (90203512)

Co-Investigator(Kenkyū-buntansha) OKABE Hiroaki  M.D., Ph.D., Kumamoto University, School of Medicine, Professor, 医学部, 教授 (20185466)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1996: ¥1,200,000 (Direct Cost: ¥1,200,000)
Keywordscapillary electrophoresis / serum protein fractions / LD isoenzymes / clinical test / M-proteimia / immunofixation electrophoresis / フリーゾーンキャピラリー電気泳動 / 生体試料微量分析
Research Abstract

Separation and quantitative estimation of the isoenzymes of lactate dehydrogenase (LD) is serum were accomplished with the Beckman P/ACE 2100 capillary electrophoresis (CE) system (Brea, CA,USA). A uncoated fused silica capillary column 50 cm long, 75 mum I.D.and substrate containing running buffer (51.6 mmol/L L-lactic acid, 8.26 mmol/L NAD in 25 mmol/L Tris (hydroxymethyl) aminoethane buffer, pH 8.7) were used. The resulting product NADH was detected at 340 nm. Injecting a sample (diluted 5 times by 25 mol/L Tris buffer, pH 8.7) by pressure 2 seconds. Separating the isoenzymes with 10kV applied for 5 min, turning off the voltage for 30 min of incubation at 24゚C and reapplying 10kV of 30 min. The results (LD1-LD5) obtained by the proposed CE method correlated well with those by REP (Helena Labs, TX.USA) gel electrophoresis system. Within-run precision CVs were excellent with 5 isoenzymes, respectively. Proposed LD isoenzymes analysis by CE system is sensitive, precise, easy for clinic … More al use. We demonstrated the separation of human serum proteins (SPE) by CE system. A 20 cm uncoated fused silica capillary column (25mum I.D.) and 150 mmol/L borate buffer (pH 10.0) as the running buffer were used. Samples were diluted 11-fold in 20 mmol/L PBS (pH 7.0) before application to CE by pressure injection for 10 second. The CE column temperature is 24゚C,a voltage of 10 kV was applied 6.5 min separating the protein fractions, the peaks were detected at 200 nm. Human serum was fractionated into approximately 10 fractions by the proposed method. The results of analysis of 100 samples using the proposed CE system correlated well with by the cellulose acetate electrophoresis method (Olympus Optimal Co., Ltd., Tokyo, Japan). Beckman multi-channeled CE system Paragon CZE 2000 was evaluated for immunofixation electrophoresis by subtraction (IFE/s). Concordance studies between IFE/s and agarose gel imunofixation electrophoresis (IFE) showed a very good agreement on 30 monoclonal gammopathy patient samples. From these results, it is concluded that IFE/s is useful and suitable tool for routine clinical laboratory. Less

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (25 results)

All Other

All Publications (25 results)

  • [Publications] Uji,Y.,et al.: "Lactate dehydrogenase isoenzymes anlysis by capillary electrophoresis" Clin Chem. 41. S236-S236 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 宇治義則、他: "キャピラリー電気泳動を用いた臨床検査法の開発" 日本臨床化学会九州支部会誌. 6. 44-55 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uji,Y.,et al.: "Separation of human proteins with capillary electrophoresis system" Proceedings of XVI international congress of clinical Chemistry. 16. 74-74 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 上野智浩、他: "キャピラリー電気泳動法によるヒト血清蛋白分画の自動化" 日本臨床検査自動化学会雑誌. 21. 327-327 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 宇治義則他: "(総説)キャピラリー電気泳動法臨床検査への応用" 日本臨床検査自動化学雑誌. 21. 755-762 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 上野智浩、他: "全自動キャピラリー電気泳動装置パラゴンCZE2000の評価" 医学検査1997. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uji,Y.,et al.: "Evaluation of Beckman Paragon CZE2000 system ; SPE and IFE/s" Clin Chem 1997. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 宇治義則、他: "キャピラリー電気泳動装置パラゴンCZE2000" Beckman Custam Care 1997. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uji, Y., et al: "Lactate dehydrogenase isoenzymes analysis by capillary electrophoresis." Clin Chem. 41. S236 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uji, Y., et al.: "Proteins and LD isoenzymes by capillary electrophoresis." Kyushu Journal of Clinical Chemistry. 6. 44-55 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uji, Y., et al.: "Separation of human proteins with capillary electrophoresis system." Proceedings of XVI International Congress of Clinical Chemistry. 16. 74 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ueno, T.et.al.: "Separation of proteins and immunofixation for subtractions by fully automated capillary electrophoresis system." Japanese Journal of Clinical Laboratory Automation. 21. 327 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uji, Y., et al.: "Capillary electrophoresis for the routine clinical laboratory." Japanese Journal of Clinical Laboratory Automation. 21. 755-762 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ueno, T., et al.: "Evaluation of fully automated capillary electrophoresis system : Paragon CZE 2000." Japanese Journal of Medical Technology. (in press.). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uji, Y.et al.: "Evaluation of Beckman Paragon CZE 2000 system. SPE and IFE/s." Clin Chem. (in press.). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uji, Y.et al.: "Beckman Paragon CZE 2000 system." Beckman Custom Care. (in press.). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 上野智浩、他: "キャピラリー電気泳動法によるヒト血清蛋白分画の自動化" 日本臨床検査自動化学会雑誌. 21. 327-327 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 宇治義則、他: "キャピラリー電気泳動法臨床検査への応用" 日本臨床検査自動化学会雑誌. 21. 755-762 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Uji Y.et al.: "Separation of human proteins with capillary electrophoresis system." Proceedings of XVI international congress of clinical chemisrty. 16. 74-74 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 上野智浩、他: "全自動キャピラリー電気泳動装置パラゴンCZE2000の評価" 医学検査. 46(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Uji Y.et al: "Evaluation of Beckman Paragon CZE2000 system ; SPE and IFE/s." Clin Chem. 43(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 宇治義則、他: "キャピラリー電気泳動装置パラゴンCZE2000" Beckman Custam CARE. 3(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Uji Y.,et.al.: "Lactatedehydrogense isoenzymes analysis by capillary electrophoresis." Clin Chem. 41. S236 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 宇治義則、他: "キャピラリー電気泳動を用いた臨床検査法の開発" 日本臨床化学会九州支部会雑誌 1996. (印刷中).

    • Related Report
      1995 Annual Research Report
  • [Publications] Uji,Y.,et.at.: "Separation of human serum proteins with capillary electrophoresis system." Anals of Clin Biochem 1996. (in press).

    • Related Report
      1995 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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