Development of a new rabbit monoclonal antibody and its based competitive indirect enzyme-linked immunosorbent assay for rapid detection of sulfonamides

J Sci Food Agric. 2013 Feb;93(3):667-73. doi: 10.1002/jsfa.5945. Epub 2012 Nov 26.

Abstract

Background: Monoclonal antibodies generally obtained through the classic mouse hybridoma system were requisite for the establishment of various immunoassays. In this study, a new rabbit monoclonal antibody (RabMAb) against sulfonamides (SAs) was first produced via hybridoma technique in rabbit. The related enzyme-linked immunosorbent assay (ELISA) was then developed and applied to real sample analysis.

Results: A sensitive competitive indirect ELISA method based on a novel RabMAb for rapid detection of sulfonamides was first established. The obtained half-maximum inhibition concentration (IC(50)) values for four SAs were all below 10 ng mL(-1) , with 0.68 ng mL(-1) sulfathiazole (STZ), 1.11 ng mL(-1) sulfadiazine (SD), 1.15 ng mL(-1) sulfapyridine (SP) and 5.27 ng mL(-1) sulfamethoxazole (SMX). Desirable recoveries when detecting different spiked swine urine and milk samples were achieved ranging from 92.6% to 104.3% and from 61.1% to 81.6%, respectively.

Conclusion: The proposed immunoassay with the newly developed RabMAb is capable of detection of four SAs (STZ, SD, SP and SMX) with proven satisfactory performance and is applicable for routine large-scale analysis in practical uses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Antibody Specificity
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Hybridomas / immunology
  • Immunization
  • Milk / chemistry
  • Rabbits
  • Sulfathiazole
  • Sulfathiazoles / immunology
  • Sulfonamides / analysis*
  • Sulfonamides / immunology*
  • Sulfonamides / urine
  • Swine

Substances

  • Antibodies, Monoclonal
  • Sulfathiazoles
  • Sulfonamides
  • Sulfathiazole