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1998 Fiscal Year Final Research Report Summary

Develepment of high sensitivity and high resolution techniques for the analysis of myeloma proteins

Research Project

Project/Area Number 09640731
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 分離・精製・検出法
Research InstitutionEhime University

Principal Investigator

MANABE Takashi  Ehime University, Faculty of Science, Department of Chemistry, Professor, 理学部, 教授 (60094281)

Co-Investigator(Kenkyū-buntansha) SHIMAZAKI Yoji  Ehime University, Fculty of Science, Assistant, 理学部, 助手 (80284389)
Project Period (FY) 1997 – 1998
Keywordsplasma proteins / capillary electrophoresis / myeloma / isoelectric focusing / SDS electrophoresis / high-sensitivity analysis / high-resolution anaiysis / two-demensional gel electrophoresis
Research Abstract

We aimed to develop techniques to analyze human plasma mycloma proteins in high sensitivity and high resolution. Microscale two-dimensional gel clectrophoresis and capillary electrophoresis were the candidates, but we found that two-dimensional gel clectrophoresis requires many steps of manual handling to obtain the separation patterns and additional processes of image analysis arc necessary to quantitate each separated proteins. Then we examined two separation principles of capillary electrophoresis, capillary isoelectric focusing and capillary SDS elcetrophoresis, for the analysis of mycloma proteins. Electrophoretic conditions for high-resolution and high-sensitivity separation of human plasma proteins have been developed for the two separation principles of capillary electrophoresis. Employing the established separation conditions, normal human plasma proteins have been separated as about 60 peaks/shoulders within 60 minutes by capillary isoelectric focusing and as about 30 peaks/shoulders within 70 minutes by capillary SDS electrophoresis. Major plasma proteins and mycloma proteins can be identified and quantitated on-column during separation by their UV absorbance. Our results show that capillary isoelectric focusing and capillary SDS electrophoresis are quite effective for high performance separation and analysis of human plasma mycloma proteins, when the separation conditions we have established were employed.

  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] 真鍋敬,宮本洋光,岩崎愛子: "Effects of Cathclytes on the mobilization of Proteins After Capillary Isoelectric Focusing" Electrophoresis. 18. 92-97 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 真鍋敬,岩崎愛子,宮本洋光: "Separation of Human Plasmal Serum Proteins by Capillary Isoelectric Focusing in the Abseme of Denaturing Agents" Electrophoresis. 18. 1159-1165 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 真鍋敬,豊田洋介,宮本洋光: "Carrier-Ampholyte-Based Separation of Basic Proteins Using Micro Twe-Dimensional Polyacrylamide Gel Electrophoresis" Chromatograpby. 18. 169-174 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 真鍋敬,宮本洋光,大田英人,豊田洋介: "Correlation of Plasma Protein Separation Patterns Obtained by Two-Dimensional Polyacylamide Gel Electrophoresis and by Capillary Electrophoresis" Electrophoresis. 19. 1319-1324 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 真鍋敬,大田英人,向井準: "Size Separation of Sodium Dodeyl Salfate Complexes of Human Plasma Proteins by Capillary Electrophoresis Employing Linear Polyacylamide as a Sieving Polywer" Electrophoresis. 19. 2308-2316 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 島崎洋次,小原清,真鍋敬: "Removal of Specific Protein Spots on the Patterus of Non-Denaturiug Two-Dimensional Electrophoresis Using ProteinA Agarose and Autibodies" J.Biochem.Biophys.Methods. 37. 1-4 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 共著者117名 (真鍋敬): "最新電気泳動実験法" 医歯薬出版(日本電気泳動学会編集) 分担計15ページ, 470 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Manabe, H.Miyamoto, A.Iwasaki: ""Effects of Catholytes on the Mobilization of Proteins After Capillary Isoelectric Focusing"" Electrophoresis. 18. 92-97 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Manabe, A.Iwasaki, H.Miyamoto: ""Separation of Human Plasma/Serum Proteins by Capillary Isoelectric Focusing in Absence of Denaturing Agents"" Electrophoresis. 18. 1159-1165 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Manabe, Y.Toyota, H.Moyamoto: ""Carrier Amopholyte-Based Separation of Basic Proteins Using Micro Two-Dimensional Polyacryamid Gel Electrophoresis"" Chromatography. 18. 169-174 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Shimazaki and T.Manabe: ""Application of two-dimensional electrophoresis without denaturing agents to comparative study of vertebrate plasma proteins"" Japanese Journal of Electrophoresis. 42. 13-18 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Manabe, H.Miyamoto, H.Oota, Y.Toyota: ""Correlation of Plasma Protein Separation Patterns Obtained by Two-Dimensional Polyacrylamide Gel Electrophoresis and by Capillary Electrophoresis"" Electrophoresis. 19. 1319-1324 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Shimazaki, T.Manabe: ""Separation of Chicken Egg Proteins Using Two-Dimensional Electrophoresis in the Absence of Denaturing Agents"" Japanese Journal of Electrophoresis. 42. 155-159 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Manabe, H.Oota, J.Mukai: ""Size Separation of Sodium Dodecyl Sulfate Complexes of Human Proteins by Capillary Electrophoresis Employing Linear Polyacrylamide As a Sieving Polymer"" Electrophoresis. 19. 2308-2316 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Shimazaki, S.Ohara, T.Manabe: ""Removal of Specific Protein Spots on the Patterns of Non-Denaturing Two-Dimensional Electrophoresis Using Protein A Agarose and Antibodies"" J.Biochem.Biophys.Methods. 37. 1-4 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Manabe, H.Mizuma, K.Watanabe: ""A Non-Denaturing Protein Map of Human Plasma Proteins Correlated With a Denaturing Polypeptide Map Combining Techniques of Micro Two-Dimensional Gel Electrophoresis"" Electrophoresis. 20 (in press).

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

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Published: 1999-12-08  

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