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稳定转化的黑腹果蝇 S2 细胞中表达和免疫原性的人结直肠癌抗原 GA733-2 与 Fc 抗体片段的重组嵌合蛋白。

Expression and immunogenicity of a recombinant chimeric protein of human colorectal cancer antigen GA733-2 and an Fc antibody fragment in stably transformed Drosophila melanogaster S2 cells.

机构信息

Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Suwon 446-701, Korea.

出版信息

Appl Biochem Biotechnol. 2010 Nov;162(5):1435-45. doi: 10.1007/s12010-010-8909-0. Epub 2010 Jan 30.

Abstract

Human colorectal cancer antigen GA733-2 fused to the immunoglobin Fc fragment (GA733-2-Fc) was expressed in stably transformed Drosophila melanogaster S2 cells, and the immunogenicity of recombinant GA733-2-Fc was investigated in mice. Recombinant GA733-2-Fc was secreted into a culture medium with a molecular mass of approximately 65 kDa. Recombinant GA733-2-Fc was purified to homogeneity using affinity fractionation with Protein A sepharose 4 Fast Flow. Recombinant GA733-2-Fc proteins elicited production of specific antibodies against recombinant GA733-2 by immunization through an intraperitoneal route. Recombinant GA733-2-Fc-induced antibodies showed a binding activity to human colorectal carcinoma HCT-116 cells. Secretory recombinant GA733-2-Fc from Drosophila S2 cell systems can be used as an effective experimental antigen for research in cancer vaccine development.

摘要

人结直肠癌抗原 GA733-2 与免疫球蛋白 Fc 片段融合(GA733-2-Fc)在稳定转化的黑腹果蝇 S2 细胞中表达,并在小鼠中研究了重组 GA733-2-Fc 的免疫原性。重组 GA733-2-Fc 分泌到具有约 65 kDa 分子量的培养基中。使用 Protein A sepharose 4 Fast Flow 进行亲和层析,将重组 GA733-2-Fc 纯化为均质。通过腹腔途径免疫,重组 GA733-2-Fc 蛋白引发针对重组 GA733-2 的特异性抗体的产生。重组 GA733-2-Fc 诱导的抗体显示与人结直肠癌细胞 HCT-116 结合活性。来自黑腹果蝇 S2 细胞系统的分泌型重组 GA733-2-Fc 可用作癌症疫苗开发研究的有效实验性抗原。

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