Comparison of three immunoassays in the screening and characterization of monoclonal antibodies against arginine-vasopressin

J Neuroimmunol. 1985 Aug;9(3-4):205-20. doi: 10.1016/s0165-5728(85)80019-9.

Abstract

A method for screening monoclonal antibodies (McAbs) to neuropeptides was evaluated using 8-arginine-vasopressin (AVP) as a model. Mice were immunized with AVP-thyroglobulin conjugate and their spleen cells were fused with X 63-Ag8.653 mouse myeloma cells. The resulting hybridoma supernatants were screened for specific antibody production using 3 different assays: solid phase enzyme radioimmunoassay in Terasaki plates (Ter-ELISA), liquid phase radioimmunoassay (LPRIA) and an immunohistochemical technique. From 2 independent fusions, 7 McAbs specific for AVP were obtained. They belonged to the IgG1 subclass and reacted more strongly to the ring part of the nonapeptide. The screening strategy proposed relies upon a crude selection of conjugate-reacting hybridomas, followed by neuropeptide-specific hybridoma identification using both LPRIA (with radioiodinated synthetic peptide) and an immunohistochemical technique (to detect natural neuropeptide). During subcloning steps Ter-ELISA is then chosen, to select for specific clones and to eliminate those reacting with the carrier thyroglobulin.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Arginine Vasopressin / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Hybridomas / immunology
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Radioimmunoassay

Substances

  • Antibodies, Monoclonal
  • Arginine Vasopressin