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Published on: July 5, 2018
[Eukaryotic expression and application of human CD226 Ig fusion protein containing 3C protease-restricted site]
Xin-Hai Zhang1, Kun Yang, Wei Jia
1Department of Immunology, Fourth Military Medical University, Xi'an 710032, China.
Aim:
To construct, express and characterize the eukaryotic expression vector of encoding human CD226 (PTA1) extracellular region Ig fusion protein gene containing 3C protease-restricted site.
Methods:
The gene fragment encoding extracellular region of human CD226 was cloned into the eukaryotic expression vector p-3C-Ig containing 3C protease-restricted site and human Ig Fc fragment gene. After sequencing, the vector was transfected into COS7 cells, and the expressed molecule was purified by affinity chromatography. Finally, the product was characterized by immunofluorescent staining and 3C protease digestion.
Results:
After expression and purification, the Ig fusion protein could bind effectively to the CD226 ligand expressed on ECV304 cells. The Fc fragment could also be cut off by 3C protease, so that the CD226 extracellular fragment was obtained.
Conclusion:
The extracellular region of human CD226 Ig fusion protein with 3C protease-restricted site is expressed successfully in COS-7 cells, which lays the foundation for the structural and functional study of this molecule.
