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Purification of the antibody against synthetic peptide from N-myc cncoprotein

Research Project

Project/Area Number 08672053
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 小児外科
Research InstitutionUniversity of Tokyo

Principal Investigator

OBANA Kazuko  Faculty of Medicine, University of Tokyo, 医学部・附属病院, 助手 (60272580)

Co-Investigator(Kenkyū-buntansha) TSUCHIDA Yoshiaki  Gumma Children's medical Center, Director, 院長 (80010164)
KAMII Yoshiyuki  Faculty of Medicine, University of Tokyo, 医学部・附属病院, 助手 (70177567)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1996: ¥1,800,000 (Direct Cost: ¥1,800,000)
KeywordsN-Myc oncoprotein / Synthetic peptide / multiple antigen peptide method / rN-Myc
Research Abstract

The importance of determining N-myc oncoprotein rather than genomic N-myc amplification has been emphasized in neuroblastoma. N-myc-specific peptides, HGRGPPTAGSTAQSPG (codon. 136-151) and GVAPPRPGGRQTSGGDH (codon. 223-239), were synthesized and injected into rabbits in conjugation with either hemocyanin or lysine core (multiple antigen peptide method). Synthesized peptides conjugated to the lysine core raised more potent antibodies than those conjugated to hemocyanin. IgG against GVAPPRPGGRQTSGGDH purified by affinity column showed a precipitation line identical to that of N-myc oncoprotein present in N-myc amplified neuroblastomas, and strongly stained the nuclei of neuroblastoma cells with N-myc amplification, while it. did not react with N-myc unamplified tumors and with c-myc/L-myc amplified tumors either with immunoblot analysis or with immunostaining. A polyclonal antibody specific for a synthetic peptide from the N-Myc oncoprotein was thus obtained.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] K.Obana,K.Hashizume: "Expression of multidrug resistance related P-glycoprotein shows good prognosis in neuroblastoma" J.Pediatr.Surg.32(3). 420-422 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Z.Hasan,K.Obana,Y.Tsuchida,et.al: "Competltive polymerase chain reaction for the determination of N myc amplification in neuroblastoma:Report of clinical cases." Eur.J.Pediatr.Surg.(in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.W.Yang,Y.Tsuchida,et.al: "Water-soluble recombinant N-Myc protein expressed in Escherichia coli." Cancer J.10. 285-290 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.W.Yang,Y.Tsutida,et.al: "Deveropment of polyclonal antibody with defined specificity against syuthetic peptides from the N-Myc oncoprotein, using multiple antigen peptide and hemocyanin conjugation methods." J.Pediatr.Surg.(in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 土田嘉昭,尾花和子,他: "神経芽腫と癌遺伝子" 医学と薬学. 37(4). 829-834 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 楊宏偉,土田嘉昭,尾花和子,他: "N-myc遺伝子産生蛋白に対するポリクローナル抗体の作成" 腫瘍マーカー研究会誌. 12. 15-17 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] A.Inoue, Z.Hasan, H.Hemmi, N.Kanda, Y.Hayashi, T.Ishizawa, Y.Tsuchida, and H.Shimatake.: "Competitive polymerase chain reaction for the quantification of N-myc gene copy number in neuroblastoma." Tumor Biol.17. 262-270 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Tsuchida, H.Hemmi, A.Inoue, K.Obana, F.Taniguchi, Y.Hayashi, N.Kanda, and H.Shimatake.: "Genetic clinical markers of human neuroblastoma with special reference to N-myc oncogene : Amplified or not amplified?" Tumor Biol.17. 65-74 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Obana and K.Hashizume: "Expression of multidrug resistance related P-glycoprotein shows good prognosis in neuroblastoma." J.Pediatr.Surg.32. 420-423 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Z.Hasan, A.Inoue, H.Ikeda, Y.Kamii, K.Obana, K.Yokomori, Y.Tsuchida, H.Hemmi and H.Shimatake.: "Competitive polymerase chain reaction for the determination of N-myc amplification in neuroblastoma : Roport of clinical cases." Eur.J.Pediatr.Surg. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.W.Yang, H.Hemmi, X.Zong, Y.Tsuchida, and H.Shimatake: "Water-soluble recombinant N-Myc protein expressed in Escherichia coli." Cancer J.10. 285-290 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.W.Yang, H.Ikeda, K.Kato, A.Nakagawara, S.H.Choi, Y.Hayashi, H.Hemmi and Y.Tsuchida.: "Deveropment of polyclonal antibody with defined specificity against synthetic peptides from the N-Myc oncoprotein, using multiple antigen peptide and hemocyanin conjugation methods." J.Pediatr.Surg.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.W.Yang, H.Hemmi, H.Ikeda, and Y.Tsuchida.: "A polyclonal antibody against synthetic peptide conserved in N-Myc protein reacts with water solible recombinant N-Myc oncoprotein." Oncol.Reports. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Tsuchida, K.Obana, et.al: "The role of subtraction of alpha-fetoprotein in the management of pediatric surgical patients" J.Pediatr.Surg.32(3). 514-517 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] K.Obana, K.Hashizume: "Expression of multidrug resistance-related P-glycoprotein shows good prognosis in neuroblastoma" J.Pediatr.Surg.32(3). 420-422 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Z.Hasan, K.Obana, Y.Tsuchida, et.al: "Competitive polymerase chain reaction for the determination of N-myc amplification in nouroblastoma:Report of clinical cases." Eur.J.Pediatr.Surg.(in-press). (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] H.W.Yang, Y.Tsuchida, et.al: "Water-soluble recombinant N-Myc protein expressed in Escherichia coli." Cancer J.10. 285-290 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 土田嘉昭, 尾花和子, 他: "神経芽腫と癌遺伝子" 医学と薬学. 37(4). 829-834 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Tsuchida,K.Obana,et al: "Genetic clinical markers of human neuroblastoma with special reference to N-myc oncogene" Tumor Biology. 17(2). 65-74 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 尾花 和子,土田,嘉昭 他: "神経芽膜と癌遺伝子" 医学と薬学. 38(3). inpress (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Tsuchida,K.Obana,et al: "The role of subfractionation of alpha-fetoprotein in the management of pediatric surgicil patients" J.Pediatr.Surg. 32(3). inpress (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Obana,et al: "Expression of multidrug resistnce-related P-glycoprotein shorus good prognosis in neuroblastoma" J.Pedictr.Surg. 32(3)(in press). (1997)

    • Related Report
      1996 Annual Research Report

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

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