Production, characterization, and applications of mouse monoclonal antibodies against gonadotropin, somatolactin, and prolactin from common carp (Cyprinus carpio)

Gen Comp Endocrinol. 2010 Jul 1;167(3):373-8. doi: 10.1016/j.ygcen.2009.10.005. Epub 2009 Oct 23.

Abstract

The gonadotropin alpha subunit (cGTH alpha), gonadotropin II beta subunit (cGTHII beta), somatolactin (cSL), and prolactin (cPRL) were isolated from the pituitaries of common carps, purified by traditional chromatographic analysis, identified by mass-chromatographic analysis, and used as immunogens in the B-lymphocyte hybridoma technique. Totally, 7, 11, 17, and 8 hybridoma cell lines were established, which were able to stably secrete monoclonal antibodies (mAbs) against cGTH alpha, cGTHII beta, cSL, and cPRL, and designated as FMU-cGTH alpha 1-7, FMU-cGTHII beta 1-11, FMU-cSL 1-17, and FMU-cPRL 1-8, respectively. The isotype, titer, and specificity were identified by enzyme-linked immunosorbent assay (ELISA), Western blot, and immunohistochemical staining, respectively, and application of these mAbs in the aforementioned tests has been proved. Furthermore, sensitive sandwich-ELISA systems for quantitative detection of the hormones mentioned above were also developed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / analysis
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / isolation & purification
  • Antibodies, Monoclonal / pharmacology*
  • Antibody Formation
  • Carps / metabolism*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Enzyme-Linked Immunosorbent Assay / standards
  • Female
  • Fish Proteins / immunology*
  • Fish Proteins / isolation & purification
  • Fish Proteins / metabolism
  • Glycoproteins / immunology*
  • Glycoproteins / isolation & purification
  • Glycoproteins / metabolism
  • Gonadotropins / immunology*
  • Gonadotropins / isolation & purification
  • Gonadotropins / metabolism
  • Immunohistochemistry / standards
  • Mice
  • Mice, Inbred BALB C
  • Pituitary Hormones / immunology*
  • Pituitary Hormones / isolation & purification
  • Pituitary Hormones / metabolism
  • Prolactin / immunology*
  • Prolactin / isolation & purification
  • Prolactin / metabolism
  • Protein Engineering / methods
  • Reference Standards

Substances

  • Antibodies, Monoclonal
  • Fish Proteins
  • Glycoproteins
  • Gonadotropins
  • Pituitary Hormones
  • somatolactin protein, fish
  • Prolactin