Simultaneous production of monoclonal antibodies against Bacillus thuringiensis (Bt) Cry1 toxins using a mixture immunization

Anal Biochem. 2017 Aug 15:531:60-66. doi: 10.1016/j.ab.2017.05.016. Epub 2017 May 17.

Abstract

The detections of Cry1 toxins are mainly dependent on immunoassays based on specific monoclonal antibodies (mAb). In the present study, a mixture immunization with seven Cry1 toxins was administered. The results showed that five mAbs with different characteristics, especially one mAb named 5-E8 which could recognize all the seven Cry1 toxins were obtained. Based on the 5-E8 mAb, a double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA) which can specifically detect the seven Cry1 toxins without cross-reactivity to Cry2A and vip3 was developed with the limit of detection (LOD) and limit of quantification (LOQ) of 6.37-11.35 ng mL-1 and 17.36-24.48 ng mL-1, respectively. The recovery tests showed that the recoveries ranged from 78% to 110% within the quantitation range (LOQ-100 ng mL-1). The established DAS-ELISA can be a useful tool for monitoring the Cry1 toxins in agricultural products. Mixture immunization opens a new path for producing diverse mAbs simultaneously in a single immunization circle.

Keywords: Cry1 family; DAS-ELISA; Diverse hybridomas; Mixture immunization; Monoclonal antibody.

MeSH terms

  • Animals
  • Antibodies, Bacterial / immunology*
  • Antibodies, Monoclonal / immunology*
  • Bacillus thuringiensis / immunology*
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / immunology*
  • Endotoxins / immunology*
  • Female
  • Hemolysin Proteins / immunology*
  • Immunization*
  • Mice
  • Mice, Inbred BALB C

Substances

  • Antibodies, Bacterial
  • Antibodies, Monoclonal
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins
  • Endotoxins
  • Hemolysin Proteins
  • insecticidal crystal protein, Bacillus Thuringiensis