Tags

Type your tag names separated by a space and hit enter

[Construction and expression of an anti-GD2,ScFv-IL-2 fusion protein gene].
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2007 Feb; 24(1):170-5.SW

Abstract

By combining interleukin2 (IL-2) with a tumor specific antibody, immunotherapy of tumors may become more effective in the future. Anti-GD2 single chain antibody directed to the extracellular domain of GD2 disialoganglioside can result in an antitumor response in some pateins with tumors expressing GD2. In this study, the fusion protein consisting of GD2 single chain antibody (ScFv) and IL-2(Ala125) was constructed. Anti-GD2 ScFv and IL-2 genes were obtained by PCR, then the ScFv-IL-2 gene was constructed by over lap PCR. The gene was inserted into the pMD18-T easy vector. Genes from pMD18-T -vector were inserted into expression vector pSE380. Recombinant expression vector was identified by restriction enzyme-cutting and then was transformed into BL21. SDS-PAGE and Western blot analysis confirmed that the transformed E. Coli BL21 could express ScFv-IL-2 fusion-proteins and the molecular weight is 43 kDa. The fusion protein was purified by affinity chromatograph and Sephacryl S-200HR then was identified through ELISA. The results show that the fusion protein retains the activities of both antigen binding and IL-2.

Authors+Show Affiliations

Group of Microbiological Engineering, Institute of Applied Ecology, Chinese academy of Sciences, Shenyang 110016, China. nijianfeng2000@126.comNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

English Abstract
Journal Article

Language

chi

PubMed ID

17333916

Citation

Ni, Jianfeng, et al. "[Construction and Expression of an anti-GD2,ScFv-IL-2 Fusion Protein Gene]." Sheng Wu Yi Xue Gong Cheng Xue Za Zhi = Journal of Biomedical Engineering = Shengwu Yixue Gongchengxue Zazhi, vol. 24, no. 1, 2007, pp. 170-5.
Ni J, Ji J, Lü A, et al. [Construction and expression of an anti-GD2,ScFv-IL-2 fusion protein gene]. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2007;24(1):170-5.
Ni, J., Ji, J., Lü, A., Huang, R., Wei, Y., & Wu, W. (2007). [Construction and expression of an anti-GD2,ScFv-IL-2 fusion protein gene]. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi = Journal of Biomedical Engineering = Shengwu Yixue Gongchengxue Zazhi, 24(1), 170-5.
Ni J, et al. [Construction and Expression of an anti-GD2,ScFv-IL-2 Fusion Protein Gene]. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2007;24(1):170-5. PubMed PMID: 17333916.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Construction and expression of an anti-GD2,ScFv-IL-2 fusion protein gene]. AU - Ni,Jianfeng, AU - Ji,Jianfei, AU - Lü,An'guo, AU - Huang,Ribo, AU - Wei,Yutuo, AU - Wu,Wenfang, PY - 2007/3/6/pubmed PY - 2009/5/22/medline PY - 2007/3/6/entrez SP - 170 EP - 5 JF - Sheng wu yi xue gong cheng xue za zhi = Journal of biomedical engineering = Shengwu yixue gongchengxue zazhi JO - Sheng Wu Yi Xue Gong Cheng Xue Za Zhi VL - 24 IS - 1 N2 - By combining interleukin2 (IL-2) with a tumor specific antibody, immunotherapy of tumors may become more effective in the future. Anti-GD2 single chain antibody directed to the extracellular domain of GD2 disialoganglioside can result in an antitumor response in some pateins with tumors expressing GD2. In this study, the fusion protein consisting of GD2 single chain antibody (ScFv) and IL-2(Ala125) was constructed. Anti-GD2 ScFv and IL-2 genes were obtained by PCR, then the ScFv-IL-2 gene was constructed by over lap PCR. The gene was inserted into the pMD18-T easy vector. Genes from pMD18-T -vector were inserted into expression vector pSE380. Recombinant expression vector was identified by restriction enzyme-cutting and then was transformed into BL21. SDS-PAGE and Western blot analysis confirmed that the transformed E. Coli BL21 could express ScFv-IL-2 fusion-proteins and the molecular weight is 43 kDa. The fusion protein was purified by affinity chromatograph and Sephacryl S-200HR then was identified through ELISA. The results show that the fusion protein retains the activities of both antigen binding and IL-2. SN - 1001-5515 UR - https://www.unboundmedicine.com/medline/citation/17333916/[Construction_and_expression_of_an_anti_GD2ScFv_IL_2_fusion_protein_gene]_ DB - PRIME DP - Unbound Medicine ER -