[Preparation and identification of PON2 monoclonal antibodies.].
OBJECTIVE
To prepare and characterize the mouse monoclonal antibodies against human PON2 (paraoxonase 2).


METHODS
A fragment of human PON2 gene of low homology with mice but of strong hydrophilicity and immunogenicity was selected for recombinant expression. Mice were immunized with the purified HIS fusion protein 3 times. The specificity and sensitivity of the anti-human PON2 monoclonal antibodies were characterized by Western blot and indirect immunofluorescence.


RESULTS
HIS-PON2 and GST-PON2 fusion protein was highly expressed in E.coli with a molecular weight of 40 kDa and 46 kDa. Western blot analysis proved that the established monoclonal antibodies could specifically recognize the target proteins expressed in E.coli expression system. Indirect immunofluorescence analysis confirmed that PON2 protein was located in the cytoplasm of HepG2 cells.


CONCLUSIONS
The monoclonal antibodies against human PON2 can specifically recognize natural protein expressed in human cells, Which can be used for further functional study of PON2 protein.
Yu
Zhan-Hong
ZH
State Key Laboratory of Proteomics, Department of Antibody Engineering, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China.
Yang
Jin-Ju
JJ
Liu
Jing
J
Chi
Jun
J
Chen
Miao
M
Liu
Xiao-Lan
XL
Liu
Ying
Y
Gao
Jian-En
JE
Sun
Qi-Hong
QH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D019084:Fluorescent Antibody Technique, Indirect; D056945:Hep G2 Cells; D006801:Humans; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins