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高等学校化学研究 ›› 2011, Vol. 27 ›› Issue (3): 441-444.

• Articles • 上一篇    下一篇

Construction, Expression and Characterization of a Chimeric Protein Targeting Carcinoembryonic Antigen in Lung Cancer

LI Yang1, HUA Shu-cheng1, MA Cheng-yuan2, YU Zhen-xiang1, XU Li-jun1, LI Dan1, SUN Li-li3, LI Xiao4 and PENG Li-ping1*   

  1. 1. Department of Respiration,
    2. Department of Neurosurgery, the First Hospital of Jilin University, Changchun 130021, P. R. China;
    3. Department of Head and Neck Surgery, Tumor Hospital of Jilin Province, Changchun 130012, P. R. China;
    4. Laboratory of Genetic Engineering of PLA, Academy of Military Medical Sciences, Changchun 130062, P. R. China
  • 收稿日期:2011-03-10 修回日期:2011-03-25 出版日期:2011-05-25 发布日期:2011-04-29
  • 通讯作者: PENG Li-ping E-mail:pengliping64@yahoo.cn
  • 基金资助:

    Supported by the Innovation Fund of Graduate of Jilin University, China(No.20101035) and the China Postdoctoral Science Foundation Project(No.20100481057).

Construction, Expression and Characterization of a Chimeric Protein Targeting Carcinoembryonic Antigen in Lung Cancer

LI Yang1, HUA Shu-cheng1, MA Cheng-yuan2, YU Zhen-xiang1, XU Li-jun1, LI Dan1, SUN Li-li3, LI Xiao4 and PENG Li-ping1*   

  1. 1. Department of Respiration,
    2. Department of Neurosurgery, the First Hospital of Jilin University, Changchun 130021, P. R. China;
    3. Department of Head and Neck Surgery, Tumor Hospital of Jilin Province, Changchun 130012, P. R. China;
    4. Laboratory of Genetic Engineering of PLA, Academy of Military Medical Sciences, Changchun 130062, P. R. China
  • Received:2011-03-10 Revised:2011-03-25 Online:2011-05-25 Published:2011-04-29
  • Contact: PENG Li-ping E-mail:pengliping64@yahoo.cn
  • Supported by:

    Supported by the Innovation Fund of Graduate of Jilin University, China(No.20101035) and the China Postdoctoral Science Foundation Project(No.20100481057).

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被引次数

2

摘要： The carcinoembryonic antigen(CEA) is an oncofetal glycoprotein known as an important clinical tumor marker and is overexpressed in several types of tumors, including colorectal and lung carcinomas. We constructed a chimeric protein that exhibits both specific binding and immune stimulating activities, by fusing staphylococcal  enterotoxin A(SEA) to the C-terminus of an anti-CEA single-chain disulfide-stabilized Fv(scdsFv) antibody     (single-chain-C-terminus/SEA, SC-C/SEA). The SC-C/SEA protein was expressed in Escherichia coli(E. coli), refolded, and purified on an immobilized Ni2+ affinity chromatography column. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and  Western blot analysis reveal that the target protein was expressed sufficiently. We used immunofluorescence assays to demonstrate that SC-C/SEA could bind specifically to human lung carcinoma cells(A549), but almost human uterine cervix cells(HeLa). We also used the L-lactate dehydrogenase(LDH) release assay to show that SC-C/SEA elicits a strong A549 tumor-specific cytotoxic T lymphocyte(CTL) response in vitro. The results suggest that SC-C/SEA shows specific activity against CEA-positive cells and has potential application in CEA-targeted cancer immunotherapy.

关键词: Carcinoembryonic antigen, Staphylococcal enterotoxin A, Single-chain disulfide-stabilized Fv(scdsFv), Escherichia coli, Anti-tumor immunity

Abstract: The carcinoembryonic antigen(CEA) is an oncofetal glycoprotein known as an important clinical tumor marker and is overexpressed in several types of tumors, including colorectal and lung carcinomas. We constructed a chimeric protein that exhibits both specific binding and immune stimulating activities, by fusing staphylococcal  enterotoxin A(SEA) to the C-terminus of an anti-CEA single-chain disulfide-stabilized Fv(scdsFv) antibody     (single-chain-C-terminus/SEA, SC-C/SEA). The SC-C/SEA protein was expressed in Escherichia coli(E. coli), refolded, and purified on an immobilized Ni2+ affinity chromatography column. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and  Western blot analysis reveal that the target protein was expressed sufficiently. We used immunofluorescence assays to demonstrate that SC-C/SEA could bind specifically to human lung carcinoma cells(A549), but almost human uterine cervix cells(HeLa). We also used the L-lactate dehydrogenase(LDH) release assay to show that SC-C/SEA elicits a strong A549 tumor-specific cytotoxic T lymphocyte(CTL) response in vitro. The results suggest that SC-C/SEA shows specific activity against CEA-positive cells and has potential application in CEA-targeted cancer immunotherapy.

Key words: Carcinoembryonic antigen, Staphylococcal enterotoxin A, Single-chain disulfide-stabilized Fv(scdsFv), Escherichia coli, Anti-tumor immunity