[Production and identification of a monoclonal antibody against human HPPCn].
OBJECTIVE
To make monoclonal antibody(mAb) against human HPPCn for the use in research on HPPCn's function and its relationship with liver diseases.


METHODS
The female BALB/c mice were immunized with the recombinant HPPCn proteins. Splenocytes and Sp2/0 cells were fused with PEG-1500. The positive clone was identified through indirect ELISA and then subcloned by limited dilution. Indirect ELISA, Western blot and Ig sub-class identification kit were used to identify the mAb's properties. By immunofluorescence experiments, we studied the cellular localization of HPPCn. The mAb epitope was also analyzed using peptide phage display technology.


RESULTS
An anti-HPPCn mAb, named W2-D5, was obtained. It belongs to IgG1 subclass. It could specially bind to human HPPCn. Furthermore, by immunofluorescence results, wo confirmed HPPCn located in the nucleus and our mAb could combined with the natural protein. With the mAb, the minimal detectable concentration was 0.1 μg/L for HPPCn; The peptide sequence of HPPCn₇₋₁₃;(IHLELRN)was identified as the epitope of the mAb.


CONCLUSIONS
An anti-HPPCn mAb with high specificity and high affinity was successfully obtained.
Huo
Jing-rui
JR
State Key Laboratory of Proteomics, Institute of Radiation Medicine, Academy of Military Medical Science, Beijing 102206, China.
Yao
Xi-ning
XN
Liu
Ying-fu
YF
Liu
Bao-cai
BC
Wu
Feng
F
Fan
Guo-cai
GC
Wang
Qing-ming
QM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology*; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D017228:Hepatocyte Growth Factor / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D017228:Hepatocyte Growth Factor