Purification of the IL-2 receptor (TAC) by ligand-affinity chromatography and utilization of the immobilized receptor for receptor-affinity chromatography (RAC) purification of IL-2, mutant IL-2, and IL-2 fusion proteins

J Invest Dermatol. 1990 Jun;94(6 Suppl):158S-163S. doi: 10.1111/1523-1747.ep12876139.

Abstract

Recombinant technology has facilitated the production of two soluble forms of human p55 interleukin-2 receptor (IL-2R) in Chinese hamster ovary cells. We have developed a ligand-affinity method for the medium-scale purification of these two soluble forms of the IL-2R, based on the biochemical interactions between the matrix-bound ligand (interleukin-2) and its soluble receptor. The affinity-purified IL-2R is further purified by anion-exchange chromatography followed by gel filtration. This method has provided enough highly pure IL-2R for structure and function studies and for use in practical applications such as high-flux drug-screening assays. The purified IL-2R subsequently has been immobilized on silica gel and employed for the purification of recombinant IL-2. Receptor-affinity-chromatography-purified IL-2 contains only a highly active monomeric form of the lymphokine, in contrast to immunoaffinity chromatography where several molecular forms of IL-2 with varying degrees of biologic activity are recovered. Receptor-affinity chromatography has been successfully applied to the purification of several mutant IL-2 as well as an IL-2-Pseudomonas exotoxin (IL2-PE40) fusion protein that is a 54.5-kDa chimeric protein in which the cell recognition domain is replaced by IL-2. The IL-2-PE40 is a potential cytotoxic agent for cells bearing the IL-2 receptor.

Publication types

  • Comparative Study

MeSH terms

  • ADP Ribose Transferases*
  • Animals
  • Bacterial Toxins
  • Chromatography, Affinity* / methods
  • Chromatography, Gel
  • Cricetinae
  • Cricetulus
  • Exotoxins / analysis
  • Female
  • Interleukin-2 / genetics
  • Interleukin-2 / isolation & purification*
  • Mutation
  • Ovary / analysis
  • Pseudomonas aeruginosa Exotoxin A
  • Receptors, Interleukin-2 / isolation & purification*
  • Recombinant Fusion Proteins / analysis*
  • Virulence Factors*

Substances

  • Bacterial Toxins
  • Exotoxins
  • Interleukin-2
  • Receptors, Interleukin-2
  • Recombinant Fusion Proteins
  • Virulence Factors
  • ADP Ribose Transferases