Expression of Recombinant Porcine Interleukin-2 and Application of Its Antibody to Immunoassays.
- Author:
In Soo CHOI
1
;
Han Sang YOO
Author Information
1. Department of Infectious Diseases, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Suwon 441-744, Korea. yoohs@plaza.snu.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
porcine IL-2;
expression;
antibody;
specificity;
species cross-reactivity
- MeSH:
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
- From:Journal of Veterinary Science
2002;3(3):207-212
- CountryRepublic of Korea
- Language:English
-
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.
