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Updated: Jul 13, 2026
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[Establishment of a NIH3T3 cell line stably expressing EGFRvIII extracellular domain and evaluation of its
Xiu-ling Jiang1, Fu-yong Bi, Hua-mao Wang
1Department of Biochemistry, Wannan Medical College, Wuhu 241001, China.
Aim:
To establish a cell line stably expressing EGFRvIIIex (epidermal growth factor receptor variant III extracellular domain) and evaluate its immunogenicity.
Methods:
NIH3T3 cells stably expressing EGFRvIIIex was obtained by screening NIH3T3 cells transfected with pLNCX2-EGFRvIIIex, a plasmid encoding EGFRvIIIex. The expression level of EGFRvIIIex was examined by immunohistochemical staining and Western blot. The cell clone with the highest expression of EGFRvIIIex named as 3T3-vIIIex was used to immunize BALB/c mice. The titer and specificity of murine antiserum was evaluated by ELISA, Western blot and immunofluorescent staining.
Results:
A NIN3T3 stable cell line with high EGFRvIIIex expression was obtained. The titer of the antiserum against human EGFRvIIIex was about 10(-5). Western blot and immunofluorescent staining analysis revealed the high specificity of the antiserum.
Conclusion:
High titer of antiserum against EGFRvIIIex can be obtained by NIH3T3 cell lines stably expressing EGFRvIIIex.
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