Optimised expression and purification of RBP4 and preparation of anti‐RBP4 monoclonal antibody

The expression level of retinol‐binding protein 4 (RBP4) protein is closely related to liver damage and plays an important role in the diagnosis and prognosis of cancer. However, the preparation of anti‐RBP4 mAb or exploration on the application of anti‐RBP4 mAb has not been reported thus far. In the present study, we constructed a pET30a‐RBP4 recombinant vector, used E. coli BL21 (DE3) as the vector to express the RBP4 recombinant protein and prepared anti‐RBP4 mAb using hybridoma technology. We performed immunohistochemical analysis on hepatocellular carcinoma (HCC) and adjacent tissues by using this anti‐RBP4 mAb. In addition to the high‐purity RBP4 recombinant protein, we successfully developed the anti‐RBP4 mAb with high affinity and specificity; it binds to natural RBP4 protein and is suitable for immunohistochemical analysis.