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Article

Construction and expression of D-dimer and GPIIb/IIIa single-chain bispecific antibody

  • Authors:
    • Zhaokui Dan
    • Zui Tan
    • Hongli Xia
    • Gan Wu
  • View Affiliations / Copyright

    Affiliations: Department of Vascular Surgery, Zhongnan Hospital of Wuhan University, Wuhan, Hubei 430071, P.R. China, Department of Hepatobiliary Gastrointestinal Surgery, Xianning Central Hospital, Xianning, Hubei 437100, P.R. China, Centre of Inspection, Detection and Testing on GMO Environmental Biosafety, The Ministry of Agriculture, Wuhan, Hubei 430062, P.R. China
  • Pages: 552-556
    |
    Published online on: May 30, 2013
       https://doi.org/10.3892/etm.2013.1132
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Abstract

The aim of this study was to construct a plasmid expressing glycoprotein IIb-IIIa (GPIIb/IIIa) and D-dimer single-chain bispecific antibody for the targeted therapy of thrombosis. The phosphorylated gene encoding the anti‑GPIIb/IIIa single-chain variable fragment (scFv) and the gene encoding the anti-D‑dimer scFv were amplified by PCR and linked in tandem by blunt-end ligation. The recombinant plasmid was transfected into the competent cell line HB2151 and identified by PCR and DNA sequencing. Then, the soluble recombinant antibody in bacterial lysates was purified by an NTA column and molecular sieve chromatography in turn. Finally, the binding specificity of the purified antibody was tested by enzyme-linked immunosorbent assay (ELISA). Results demonstrated that the construction of the expression plasmid was successful and the purified recombinant protein, which had a molecular weight of ~56 kDa, was specific to GPⅡb/Ⅲa and D-dimer. In conclusion, a plasmid expressing a bispecific antibody was constructed by a new method of blunt‑end ligation. The soluble recombinant protein is a promising platform for target-oriented thrombolytic therapy.
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Copy and paste a formatted citation
Spandidos Publications style
Dan Z, Tan Z, Xia H and Wu G: Construction and expression of D-dimer and GPIIb/IIIa single-chain bispecific antibody. Exp Ther Med 6: 552-556, 2013.
APA
Dan, Z., Tan, Z., Xia, H., & Wu, G. (2013). Construction and expression of D-dimer and GPIIb/IIIa single-chain bispecific antibody. Experimental and Therapeutic Medicine, 6, 552-556. https://doi.org/10.3892/etm.2013.1132
MLA
Dan, Z., Tan, Z., Xia, H., Wu, G."Construction and expression of D-dimer and GPIIb/IIIa single-chain bispecific antibody". Experimental and Therapeutic Medicine 6.2 (2013): 552-556.
Chicago
Dan, Z., Tan, Z., Xia, H., Wu, G."Construction and expression of D-dimer and GPIIb/IIIa single-chain bispecific antibody". Experimental and Therapeutic Medicine 6, no. 2 (2013): 552-556. https://doi.org/10.3892/etm.2013.1132
Copy and paste a formatted citation
x
Spandidos Publications style
Dan Z, Tan Z, Xia H and Wu G: Construction and expression of D-dimer and GPIIb/IIIa single-chain bispecific antibody. Exp Ther Med 6: 552-556, 2013.
APA
Dan, Z., Tan, Z., Xia, H., & Wu, G. (2013). Construction and expression of D-dimer and GPIIb/IIIa single-chain bispecific antibody. Experimental and Therapeutic Medicine, 6, 552-556. https://doi.org/10.3892/etm.2013.1132
MLA
Dan, Z., Tan, Z., Xia, H., Wu, G."Construction and expression of D-dimer and GPIIb/IIIa single-chain bispecific antibody". Experimental and Therapeutic Medicine 6.2 (2013): 552-556.
Chicago
Dan, Z., Tan, Z., Xia, H., Wu, G."Construction and expression of D-dimer and GPIIb/IIIa single-chain bispecific antibody". Experimental and Therapeutic Medicine 6, no. 2 (2013): 552-556. https://doi.org/10.3892/etm.2013.1132
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