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

FEBS Open Bio. 2022 Feb;12(2):430-442. doi: 10.1002/2211-5463.13349. Epub 2021 Dec 24.

Abstract

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.

Keywords: RBP4; immunohistochemical analysis; monoclonal antibody; recombinant protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal
  • Carcinoma, Hepatocellular* / genetics
  • Escherichia coli / metabolism
  • Humans
  • Liver Neoplasms* / genetics
  • Recombinant Proteins / genetics
  • Retinol-Binding Proteins, Plasma / genetics
  • Retinol-Binding Proteins, Plasma / metabolism

Substances

  • Antibodies, Monoclonal
  • RBP4 protein, human
  • Recombinant Proteins
  • Retinol-Binding Proteins, Plasma