[High-level expression of 6B11 ovarian carcinoma anti-idiotypic antibody scFv genes in E. Coli].
OBJECTIVE
To express high-level 6B11 ovarian carcinoma anti-idiotypic antibody single chain Fv (scFv) genes in E. Coli.


METHODS
Using PCR cloning technique, We cloned 6B11scFv genes into bacterial expression vector pKPL-3a. Recombinant plasmid vector pL-6B11scFv was transformed into E. Coli pop2136 with temperature control to produce proteins. Renaturated proteins were purified on DEAE-Sepharose Fast Flow ion exchange column with salt gradient elution. Immunoactivity was determined with ELISA and inhibition ELISA tests respectively.


RESULTS
6B11scFv genes were expressed as inclusion bodies in the cytoplasm. The purity of 6B11scFv turned out by SDS-PAGE was more than 95%. The expressed proteins after purification specifically reacted with anti-ovarian carcinoma monoclonal antibody COC166-9 and efficiently inhibited the reaction between primary ovarian carcinoma antigen OC166-9 and COC166-9.


CONCLUSIONS
We successfully expressed high-level 6B11scFv genes in E. Coli. Expressed proteins showed pretty good immunoactivity.
Liu
Y
Y
Gynecologic Oncology Center, People's Hospital Beijing Medical University, Beijing 100034.
Qian
H
H
Huang
H
H
D000888:Antibodies, Anti-Idiotypic / Q000096:biosynthesis* / Q000235:genetics; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D010051:Ovarian Neoplasms / Q000276:immunology*; D016133:Polymerase Chain Reaction
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm