Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography

Mol Med Rep. 2017 Jun;15(6):4035-4040. doi: 10.3892/mmr.2017.6523. Epub 2017 Apr 27.


The immunoglobulin superfamily member CD147 is a widely expressed glycoprotein that occurs in both a membrane‑spanning and soluble form. Sandwich ELISA is a powerful tool for analyzing soluble antigens. The aim of the present study was to obtain a highly specific polyclonal antibody against human CD147 that can be used for sandwich ELISA analysis. Expression of recombinant CD147 by a eukaryotic expression system was used to immunize rabbits to obtain antiserum. A highly specific polyclonal antibody that was able to detect soluble CD147 in sandwich ELISA was obtained by antigen‑immunoaffinity chromatography purification. The purity of rabbit anti‑CD147 polyclonal antibodies was ~99%, and ELISA analysis was able to determine the titer of the rabbit anti‑CD147 polyclonal antibodies at 1:512,000. The lowest concentration of the standard CD147 antigen that the sandwich ELISA was able to detect was 31.25 pg/ml. The sandwich ELISA system was composed of anti‑hepatoma HAb18 monoclonal antibodies and purified rabbit anti‑CD147 polyclonal antibodies. The present study demonstrated that antigen‑immunoaffinity chromatography may be a good technique for the purification of polyclonal antibodies, which may be used to detect antigen in sandwich ELISAs.

MeSH terms

  • Animals
  • Antibodies / immunology*
  • Antibodies / isolation & purification*
  • Basigin / immunology*
  • CHO Cells
  • Chromatography, Affinity* / methods
  • Cricetulus
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay*
  • Humans
  • Immune Sera
  • Immunoglobulin G / immunology
  • Immunoglobulin G / isolation & purification
  • Rabbits
  • Recombinant Proteins / immunology
  • Sensitivity and Specificity


  • Antibodies
  • Immune Sera
  • Immunoglobulin G
  • Recombinant Proteins
  • Basigin