[Establishment of a NIH3T3 cell line stably expressing EGFRvIII extracellular domain and evaluation of its immunogenicity].
OBJECTIVE
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.


CONCLUSIONS
High titer of antiserum against EGFRvIIIex can be obtained by NIH3T3 cell lines stably expressing EGFRvIIIex.
Jiang
Xiu-ling
XL
Department of Biochemistry, Wannan Medical College, Wuhu 241001, China.
Bi
Fu-yong
FY
Wang
Hua-mao
HM
Shi
Bi-zhi
BZ
Zhang
Jie
J
Wang
Ming
M
Li
Zong-hai
ZH
D000818:Animals; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D066246:ErbB Receptors / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D007106:Immune Sera / Q000276:immunology; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D041681:NIH 3T3 Cells / Q000276:immunology* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D014162:Transfection
D007106:Immune Sera; C410723:epidermal growth factor receptor VIII; D066246:ErbB Receptors