Affiliations 

  • 1 State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Zhejiang-Malaysia Joint Research Laboratory for Agricultural Product Processing and Nutrition and College of Food Science and Engineering, Ningbo University, Ningbo 315800, China
  • 2 State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Zhejiang-Malaysia Joint Research Laboratory for Agricultural Product Processing and Nutrition and College of Food Science and Engineering, Ningbo University, Ningbo 315800, China. Electronic address: lanhangzhen@nbu.edu.cn
  • 3 College of Life Science, Nanjing Normal University, Nanjing, Jiangsu 210023, China
  • 4 Zhoushan Institute for Food and Drug Control, Zhoushan 316012, China
Enzyme Microb Technol, 2024 Sep 04;181:110507.
PMID: 39241682 DOI: 10.1016/j.enzmictec.2024.110507

Abstract

An immunoassay method based on penicillin-binding protein (PBP) was developed for the quantitative determination of 10 kinds of beta-lactam antibiotics (BLAs). First, two kinds of PBPs, which are named PBP1a and PBP2x, were expressed and purified, and they were characterized by SDS-PAGE and western blotting analysis. Then, the binding activity of PBP1a and PBP2x to template BLAs, cefquinome (CEFQ) and ampicillin (AMP), was determined. The effect of the buffer solution system, e.g., pH, ion concentration, and organic solvent, on the immune interaction efficiency between PBPs and BLAs was also evaluated. In the end, the PBP-based immunoassay method was developed and validated for the detection of 10 kinds of BLAs. Under optimal conditions, PBPs exhibited high binding affinity to BLAs. In addition, this method showed a high sensitivity for the detection of 10 kinds of BLAs with the limits of detection from 0.21 to 9.12 ng/mL, which are much lower than their corresponding maximum residual limit of European Union (4-100 ng/mL). Moreover, the developed PBP-immunoassay was employed for BLA detection from milk samples, and satisfactory recoveries (68.9-101.3 %) were obtained.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.