Development of a Monoclonal Antibody-Based Sandwich ELISA for Detection of Guinea Pig Interleukin-2

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概要

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• Interleukin 2 (IL-2) is a T cell proliferation factor released by Th0- and Th1-type helper T cells and is an essential cytokine for immune responses. In the present study, recombinant glutathione S-transferase (GST)-guinea pig IL-2 (GPIL-2) fusion protein was prepared by Escherichia coli (E. coli) and by using this protein as an immunogen, monoclonal antibodies (mAbs) against GPIL-2 were produced to establish a basis for a research on immune responses in guinea pigs. Three stable hybridoma cell lines were established, and specific binding of each mAb to recombinant GPIL-2 produced by E. coli and insect cells infected with recombinant baculovirus was shown by enzyme linked immunosorbent assay (ELISA) and/or immunoblot analysis. Isotype analyses of these mAbs revealed that all three mAbs were IgG1 and had κ chain. Furthermore, assessment of their epitopes by competition binding assay indicated that the mAbs obtained in this study bound to three different epitopes. Thus, a sandwich ELISA based on the two mAbs specific to different GPIL-2 epitopes was developed for detection of GPIL-2, which had a sensitivity threshold of about 0.3 ng/ml of GPIL-2.

• 公益社団法人 日本獣医学会の論文

著者

• Kato Hirotomo

  Department Of Microbiology And Oral Biology Immunobiology Vaccine Center University Of Alabama At Bi

• Takeyoshi Masahiro

  Hita Laboratory Chemicals Evaluation And Research Institute (ceri)

• Adachi Satoshi

  Department Of Energy Sciences Tokyo Institute Of Technology

• Hashimoto Tomomi

  Department Of Applied Chemistry Graduate School Of Science And Engineering Ehime University

• IWATA HIROYUKI

  Department of Applied Chemistry, Graduate School of Engineering, Tokyo Metropolitan University

• Iwata Hiroyuki

  Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University

• HASHIMOTO Tomomi

  Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University

• TAKEYOSHI Masahiro

  Hita Laboratory, Chemicals Evaluation and Research Institute

• ADACHI Satoshi

  Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University

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