J Vet Sci.  2002 Sep;3(3):207-212.

Expression of Recombinant Porcine Interleukin-2 and Application of Its Antibody to Immunoassays

• Choi IS
• Yoo HS

Abstract

Interleukin-2 plays an important role in T lymphocyte proliferation and immune response regulations. In this study, porcine IL-2 cDNA was cloned from peripheral blood mononuclear cells, and recombinant porcine IL-2 (rpIL-2) was expressed in Escherichia coli. The size of rpIL-2 without signal peptides was about 15 kDa when determined by SDS-PAGE and Western blotting analysis. Anti-rpIL-2 antibody was produced from mice immunized with the purified rpIL-2, and its specificity was examined by Western blotting and ELISA. In the Western blotting assay, anti-rpIL-2 and anti-recombinant human IL-2 (rhIL-2) antibodies specifically recognized rpIL-2 and rhIL-2, respectively. However, anti-rpIL-2 antibody did not recognize rhIL-2, and anti-rhIL-2 antibody also did not react with rpIL-2 in the same assay. In ELISA, anti-rpIL-2 antibody strongly interacted with both rpIL-2 and rhIL-2, and anti-rhIL-2 antibody also efficiently recognized both proteins. Taken together, the specificity of anti-rpIL-2 antibody for rpIL-2 was demonstrated by Western blotting and ELISA. It was also shown that ELISA is more efficient than Western blotting in determining the species cross-reactivity of anti-rpIL-2 antibody.

Keyword

porcine IL-2; expression; antibody; specificity; species cross-reactivity

MeSH Terms

Animals
Antibodies/diagnostic use/*metabolism
Antibody Specificity
Blotting, Western/veterinary
Cloning, Molecular
Cross Reactions
Electrophoresis, Polyacrylamide Gel/veterinary
Enzyme-Linked Immunosorbent Assay/*methods/veterinary
Escherichia coli/genetics/metabolism
Humans
Interleukin-2/biosynthesis/genetics/*immunology
Mice
Recombinant Proteins/biosynthesis/genetics/immunology
Species Specificity
Swine