Objective    To screen human anti-hypoxia-inducible factor (HIF)-1α scFv of lung adenocarcinoma from large phage antibody library and identify the positive clones.     Methods    Panning of large phage antibody library against lung adenocarcinoma cell line A549 and HIF-1α was conducted respectively to select specific antibodies. E. coli HB2151 was infected to induce the expression of soluble scFv. The binding activity and specificity were tested by ELISA and immunocytochemical assay. The expression and relative molecular weight of the soluble scFv was detected by SDS-PAGE and Western blot analysis.     Results    After panning

the target scFv was enriched

and ELISA resu

lts showed that positive reactions to HIF-1α were detected in 5 of 10 random clones with a positive ratio of 50%. Immunocytochemical analysis showed the specific affinity of the antibodies to A549 cells. The soluble human anti-HIF-1α scFv fragments of lung adenocarcinoma were expressed in E. coli HB2151 and then confirmed by SDS-PAGE. The result of Western blotting showed that the relative molecular weight of the soluble scFv was about 30×10

3

. The binding activity and specificity were confirmed by ELISA.     Conclusion    Human anti-HIF-1α scFv of lung adenocarcinoma is successfully obtained with large phage antibody library technique.