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Dvelopment of microanalysis for biological substances detected by bioluminescent assay

Research Project

Project/Area Number 09557219
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Laboratory medicine
Research InstitutionShowa University

Principal Investigator

MAEDA Masako  School of Pharmaceutical Sciences, Showa University, Professor, 薬学部, 教授 (00053869)

Co-Investigator(Kenkyū-buntansha) KOKADO Amane  School of Pharmaceutical Sciences, Showa University, Assistant, 薬学部, 助手 (20266159)
ITO Katsutoshi  School of Pharmaceutical Sciences, Showa University, Assistant Professor, 薬学部, 講師 (20223141)
ARAKAWA Hidetoshi  School of Pharmaceutical Sciences, Showa University, Associate Professor, 薬学部, 助教授 (70129807)
TAJIMA Noriko  School of Pharmaceutical Sciences, Showa University, Assistant, 薬学部, 助手 (30297022)
金光 まひな (保坂 まひな)  昭和大学, 薬学部, 助手 (30286869)
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥11,000,000 (Direct Cost: ¥11,000,000)
Fiscal Year 1999: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1998: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1997: ¥5,400,000 (Direct Cost: ¥5,400,000)
KeywordsBioluminescence / luciferase / Biological Substances / Enzyme Immunoassay / Analysis of Gene / Photo imaging / Acetate Kinase / Pyruvate Phosphate Dikinase / 生物発光法 / 生体成分 / BL-EIA / 診断への応用 / ブロッティングアッセイ / 制限酵素断片長多型 / BLEIA / 酵素イムノアッセイ / TSH / ヒト生長ホルモン / インターロイキン
Research Abstract

Recently, bioluminescence is extensively investigated by several researchers for its high sensitivity with the aim of applying several assays. And the various luciferases and photoprotein genes were cloned and expressed in E.coli or saccharomyces serevisiae. Especially, recombinant firefly luciferase is now commercially available and has been employed for enzyme immunoassay(EIA). However, the firefly luciferase tends to lose much of its activity when treated with the chemical cross linkage reagents used in the conjugation. To dissolve this problem, we have established two highly sensitive bioluminescent assays for thermostable acetate kinase (AK). After EIA using AK as label enzyme, AK activity was determined by measuring the amount of produced AT P using firefly luciferase.
Furthermore, we have introduced well known streptavidin-biotin system to EIA using biotinyted AK. Using these systems, we applied to assay for biologically substances such as hCG, mIL-6 and other clinically importan … More t substances. Especially, in the TSH assay, sensitivity was about 25 times higher than that of RIA. It will be useful for detection of the low concentration of TSH values such as hyperthyroidism. It have been applied to BLEIA in combination with the streptavidin-biotin system with biotinylated AK and appeared sensitive, simple, and convenient. Using the streptavidin-biotin system, mIL-6 in mouse tissue and plasma, and PACAP 38 in rat tissues: it could be accurately measured without the concentration of samples or any other pretreatment using the BLEIA. More recently, Kajiyama et al conducted mutational analysis of firefly luciferase and showed that they emit light of different colors due to single amino acid change. The study demonstrated multiple simultaneous antigen detection using the two kinds of biotinylated luciferase-streptavidin-bioanylated antibody complexes that were capable of emitting difference colors of light. As clinical application using this system, PGI and PGII in serum can be assayed, satisfactory. Further investigation is now ongoing to determine if multiple analyte in the same sample can be assayed by BLEIA. Less

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (39 results)

All Other

All Publications (39 results)

  • [Publications] M. Maeda 他: "TR-FIA for PACAP"Biomedical Analysis. 15. 1489-1495 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Maeda 他: "Bl-E1A using Thermostable Mutant Luciferase and acetate kinase as a Label Enzyme"Anal. Chim Acta. 361. 19-26 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Maeda 他: "Biocheminescent E1A with Biotinylated Firefly Luciferase"J. Clin. Ligand Assay. 21. 358-362 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Maeda 他: "BLEIA for PACAP38"Anal. Sci.. 382. 245-251 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Maeda 他: "Bioluminascent PCR-RFLP ELISA for analysis of Vitamin D receptor Gene Polymorphism"Anal. Sci.. 15. 943-949 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Maeda 他: "Simultaneous assay of pepsinogen I and pepsinogenII in serum by BLE1A using two kinds of luciola lateralis luciferase"Anal Chim. Acta.. 382. 262-272 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 前田昌子: "イムノアッセイ<免疫化学的測定法>"日本分析化学会. 30 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 前田昌子: "大学と科学<先端材料の魔術師>"文部省. 290(125-135) (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A.Kokado, A.Tsuji and M.Maeda: "Chemiluminescence assay of alkaline phosphatase using cortisol-21-phosphate as substrate and its application to enzyme immunoassay"Anal. Chim. Acta. 337. 335-340 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S.Murakami, K.Ito, T.Goto, S Kamada and M.Maeda: "Bioluminescent enzyme immunoassay using thermostable mutant luciferase and acetate kinase as a label enzyme"Anal. Chim. Acta. 361. 19-26 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Fukuda, H. Tatsumi and M. Maeda: "Bioluminescent enzyme immunoassay with biotinylated firefly luciferase"J. Clin. Ligand Assay. 21. 358-362 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Ito, T.Goto S Murakami and M.Maeda: "Bioluminescent enzyme immunoassay for mouse interleukin-6 using acetate kinase as"Anal. Sci.. 15. 91-93 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Ito, T.Goto, S Murakami, M. Maeda, S. Shiota and A. Arimuga: "Bioluminescent enzyme immnoassay for pituitary adenylate cyclase activating polypeptaide 38 (PACAP38)"Anal. Chim. Acta. 382. 245-251 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H. Arakawa, A. Kokado, S.Yoshizawa, M.Maeda, A. Tokita and Y. Yamashiro: "Bioluminescent PCR-RFLP enzyme-linked immunosorbent assay for analysis of vitamin D receptor gene polymorphism"Anal. Sci.. 15. 943-949 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H. Ohkuma, K. Abe, Y. Kosaka and M.Maeda: "Simultaneous assay of pepsinogen I and pepsinogen II in serum by bioluminescent enzyme immunoassay using two kinds of luciola lateralis luciferase"Anal. Chim. Acta. 382. 262-272 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H. Ohkuma, K.Abe, Y. Kosaka, M.Maeda: "Detection of luciferase having two kinds of luminescent colour based on optical filter procedure : application to an enzyme immunoassay"Luminescence. 15. 21-27 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] MASAKO MAEDA: "Biolumi nescent EIA for PACAB38."Anal.Chim Acta. 382. 245-251 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] MASAKO MAEDA: "Biolumi neycent PCR-RFLP-ELISA for Analysis of Vitamine D Receptor Gane Poly mor khism"Anal.Sci.. 15. 943-949 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] MASAKO MAEDA: "Simultaneous asyay of pepsinogenl and 11 in serum by Biolumi gres cent EIA using 2kinds of luciferase"Anal.Chim Acta. 395. 265-272 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] MASAKO MAEDA: "Themi lumi nes cence assay for catecholanime by the generation of H_2O_2 in pasic solution and use of luniorol."Anal.Sci.. 15. 1269-1272 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] MASAKO MAEDA: "Development of Novel High Sensitinity chevi luminescence Assoy for Lumi nol Uesing this urea derivotiones"Lumines cence.. 14. 355-360 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] MASAKO MAEDA: "Chemi lumines cent Determination of Lucigenin using this ureid Derivatiues."Anal.chem Acta. 403. 125-130 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] 前田 昌子: "分析化学ワークブック"南江堂(分担). 180 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] MASAKO MAEDA: "Bioluminescent Engyme lmmunoassay using thermostable mutant Luayeraos and Acetate Kinase as a Label Engyne" Anal.Chim.Acta. 361. 19-26 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] MASAKO MAEDA: "Developement of on EIA for antitumon agent MKT077" Anal.Chim.Acta. 362. 177-186 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] MASAKO MAEDA: "Analysis of PCR Products by HPLC with Fluovimentic Deteetion and its Application to DNA diagnosis" J.chromatogr. A.787. 261-265 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] MASAKO MAEDA: "Chemilumines cent Assay of β-o-Galaeto sidase Basecp on Jndole Luminescence" J.Brolumin.Chemilumin.13. 349-354 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] MASAKO MAEDA: "Monoclonal-based ELISA and lmmunochromato graphre Rapid Assay for Monensin." The Analyst.123. 2573-2578 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] MASAKO MAEDA: "Bioluminescent EIA of mouse lnterlenkin 6 using Acetate Kinase" Analytical Sciences. 15. 91-93 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] 前田 昌子: "基礎薬学,分析化学I" 広川書店, 380 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 前田 昌子: "実践臨床検査医学" 文光堂, 490 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] MASAKO MAEDA: "Cheeniluminescence assay of Alp.using Cortisol-21-p as suustrate and its application to enzyme immunoassay" Anal Chim Acta. 337. 335-340 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] MASAKO MAEDA: "Cheniluminescence FIA of Alp and its application to EIA" Anal.Chim Acta. 339. 139-146 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] MASAKO MAEDA: "Constrnction of Time・Resolned Fluores,Detection for Amino Compounds after HPLC using Europium Chelate." Anal.Chem.69. 1861-1865 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] MASAKO MAEDA.: "Time Resolned FIA for PACAP using Eu (III) ion chelate Labeled STV-Biotin Conyslex" J.pharm.Biomed.Analysis.15. 1489-1495 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] MASAKO MAEDA: "Enranced Resolution in the Capillany Electrophoresis Separation of Puble-stranded DNA" Anal.Sci. 13. 853-855 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] MAEDA MASAKO: "Indirect TR-FIAetection of both Non-fluorescent and Fluorescent Compounds Seporated by HPLC" J.chromatogs.A. 787. 261-265 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 前田 昌子: "イムノアッセイ〈免疫化学的測定〉" 日本分析化学会(ビデオ 30分), (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 前田 昌子: "分析化学II" 広川書店, 400 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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