Affiliation:
1. Henan Institute of Science and Technology, College of Animal Science and Veterinary Medicine, Xinxiang 453003, China
Abstract
The purpose of this experiment was to study the design and modification of hapten molecules and artificial antigen molecules of deoxynivalenol (DON), and to compare the preparation and identification methods of four artificial antigens. According to the characteristics of the molecular structure of DON, four artificial antigen coupling methods were designed—namely, N,N′-carbonyldiimidazole (CDI), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), isobutyl chloroformate (IBCF), and N-hydroxysuccinimide (NHS)—to prepare artificial antigens and detection antigens. Through ultraviolet (UV), infrared (IR), and SDS–polyacrylamide gel electrophoresis (SDS–PAGE), along with other physical and chemical identification methods and animal immunisation, the best artificial antigen coupling method was screened. The results showed that the CDI method achieved the best effect among the synthesis methods. The titre of anti-DON polyclonal antibody (pAb) produced by animal immunisation reached 1: (6.4 × 103). The half inhibitory concentration (IC50) was 47.75 ng/mL, the cross-reaction rate with 3-acetyldeoxynivalenol (3-AcDON) was slightly higher at 35.3%, and there was no cross-reaction with other compounds; therefore, four artificial antigens were successfully prepared by using the molecular structure of DON. Through identification, the CDI method was screened as the best artificial antigen synthesis method, with the highest DON pAb titre, the best sensitivity, and the strongest specificity. This will lay a solid antigenic foundation for the preparation of better anti-DON monoclonal antibodies (mAbs) in the future.
Funder
Key R&D and promotion projects in Henan Province in 2023
introduction of leading talents and the construction of high-level talents team of Henan Institute of Science and Technology
2022 innovation and entrepreneurship training program for College Students of Henan Institute of Science and Technology
2023 Key Scientific Research Project of Universities in Henan Province
Twelfth Five-Year Plan of National Science and Technology Support Projects, “Research and Demonstration of Rapid Detection Technology for Hormone Drugs”
Program for Innovative Research Team (in Science and Technology) in University of Henan Province
Central Plains Science and Technology Innovation Leading Talent Project
Subject
Chemistry (miscellaneous),Analytical Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Molecular Medicine,Drug Discovery,Pharmaceutical Science