Preparation and identification of monoclonal antibody against abrin-a

J Agric Food Chem. 2011 Sep 28;59(18):9796-9. doi: 10.1021/jf202534y. Epub 2011 Sep 6.

Abstract

BALB/c mice were immunized four times with formalin-prepared abrin-a. Using the polyethylene glycol method, immunized splenocytes were isolated and fused with SP2/0 cells. An indirect ELISA was established and used to detect positive clones secreting monoclonal antibodies (mAbs) against abrin-a. After analysis, three hybridoma clones secreting IgG-subtype mAbs were obtained. The antibodies were purified from the hybridoma growth medium using protein A or G affinity chromatography. Western blot analysis was used to analyze the antigenic epitopes on abrin-a recognized by the mAbs. The mAbs were specific for abrin-a, with no detectable cross-reactivity with several homologous toxins and associated agglutinins. Sandwich ELISA was then developed using these mAbs, which had a detection limit for abrin-a of 7.8 ng/mL.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abrin / analysis
  • Abrin / immunology*
  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / isolation & purification
  • Antibody Specificity
  • Culture Media, Conditioned / chemistry
  • Enzyme-Linked Immunosorbent Assay / methods
  • Female
  • Hybridomas / immunology
  • Immunization
  • Immunoglobulin G / immunology
  • Immunoglobulin G / isolation & purification
  • Mice
  • Mice, Inbred BALB C

Substances

  • Antibodies, Monoclonal
  • Culture Media, Conditioned
  • Immunoglobulin G
  • Abrin