Korean Journal of Fisheries and Aquatic Sciences (한국수산과학회지)

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Production of Monoclonal Antibodies Against the Immunoglobulin M of Olive Flounder Paralichthys Olivaceus

넙치(Paralichthys olivaceus)의 immunoglobulin M에 대한 단클론 항체 생산

  • Kim, Wi-Sik (Department of Aqualife Medicine, Chonnam National University) ;
  • Kim, Ki-Hong (Department of Aqualife Medicine, Chonnam National University) ;
  • Kim, Choon-sup (EnbioGene) ;
  • Oh, Myung-Joo (Department of Aqualife Medicine, Chonnam National University)
  • 김위식 (전남대학교 수산생명의학과) ;
  • 김기홍 (전남대학교 수산생명의학과) ;
  • 김춘섭 ((주)엔바이로젠) ;
  • 오명주 (전남대학교 수산생명의학과)
  • Received : 2017.02.10
  • Accepted : 2017.03.23
  • Published : 2017.04.30
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Abstract

Immunoglobulin M (IgM) was purified from olive flounder Paralichthys olivaceus sera using mannan-binding protein (MBP) and protein L affinity columns (designated as MBPIgM and ProLIgM, respectively). A monoclonal antibody (MAb) against olive flounder IgM was produced. The MBPIgM and ProLIgM had apparent molecular weights of 77, 73, and 28 kDa in SDS-PAGE. Nine hybridomas secreting MAbs against olive flounder IgM were established: five MAbs for MBPIgM (1, 2, 3, 4, and 5) and four for ProLIgM (6, 7, 8, and 9). Western blotting indicated that seven MAbs recognized heavy (H; MAbs 1, 2, 3, 4, 5, 6, and 7) chains and one recognized light (L; MAb 9) chains of IgM, while MAb 8 did not recognize IgM. The results of enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the nine MAbs revealed that the optical density (OD) values of sera differed significantly between BSA- and non-immunized fish, despite some sera from non-immunized fish with slight high OD values. These results suggest that the MAbs produced in this study reacted specifically with the IgM from olive flounder.

Keywords

References

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