[Expression and purification of T7 bacteriophage capsid protein P11 and preparation of monoclonal antibody against P11].
OBJECTIVE
To express capsid tail protein P11 of T7 bacteriophage and produce mouse monoclonal antibody(mAb) against the protein.


METHODS
P11 protein was cloned and recombinant P11 protein was expressed as a fusion protein with an N-terminal 6-His tag. The purified protein was used to immunize BALB/c mouse. The specificity of mAb was analyzed by ELISA and Western blot.


RESULTS
P11 protein was successfully expressed and purified. SDS-PAGE analysis showed that the molecular weight of the expressed protein was approximately 27 kd. One hybridoma cell (2G11) secreting mAb against P11 was developed. The isotype of the mAb was IgG(2b). ELISA detection showed that titers of mAb was 1:8. 1 x 10(5) in ascites.Western blot analysis proved mAb obtained could react specifically to the recombinant p11 protein.


CONCLUSIONS
Recombinant P11 protein and mAbs were successfully prepared.
Chen
Liang
L
College of Veterinary Medicine, Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100, China. cl198211@yahoo.com.cn
Gao
Yun-ying
YY
Wang
Hua-mao
HM
Xu
Rong
R
Liu
Su-xia
SX
Li
Zong-hai
ZH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D017123:Bacteriophage T7 / Q000737:chemistry* / Q000235:genetics; D036022:Capsid Proteins / Q000276:immunology*; D002459:Cell Fusion; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D014764:Viral Proteins / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D007074:Immunoglobulin G; D014764:Viral Proteins