TAILIEUCHUNG - Báo cáo khoa học: " Expression of Recombinant Porcine Interleukin-2 and Application of Its Antibody to Immunoassays"

Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Expression of Recombinant Porcine Interleukin-2 and Application of Its Antibody to Immunoassays | J. Vet. Sci. 2002 3 3 207-212 JOURNAL OF Veterinary Science Expression of Recombinant Porcine Interleukin-2 and Application of Its Antibody to Immunoassays In-Soo Choi and Han-Sang Yoo Department of Infectious Diseases College of Veterinary Medicine and School of Agricultural Biotechnology Seoul National University Suwon 441-744 Korea Received February. 8 2002 Accepted July 20 2002 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. Key words porcine IL-2 expression antibody specificity species cross-reactivity Introduction Interleukin-2 IL-2 was originally identified as a T cell growth factor in the supernatants of lectin-stimulated human and mouse T lymphocytes 11 21 26 . IL-2 plays a number of important roles in the regulation of immune responses including the proliferation of activated T lymphocytes the generation of cytotoxic T .

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