Production of Natural Penicillin-Binding Protein 2 and Development of a Signal-Amplified Fluorescence Polarization Assay for the Determination of 28 Beta-Lactam Antibiotics in Milk

被引:1
作者
Hu, Jia Jia [1 ]
Wang, Jian Ping [1 ]
机构
[1] Hebei Agr Univ, Coll Vet Med, Baoding 071000, Hebei, Peoples R China
基金
中国国家自然科学基金;
关键词
beta-lactam antibiotics; magnetic activity-based proteinprofiling probe; penicillin-binding protein 2; signalamplification fluorescence polarization assay; recognitionmechanism; STAPHYLOCOCCUS-AUREUS; IMMUNOASSAYS; INHIBITION; BLAR;
D O I
10.1021/acs.jafc.4c07028
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
In this study, two magnetic activity-based protein profiling probes based on cephradine and amoxicillin were first synthesized that were used to produce the natural penicillin-binding protein 2 of Escherichia coli. After characterization by using LC-ESI-MS/MS, it was found that the obtained proteins by using the two probes were the same. The molecular docking for 28 beta-lactam antibiotics showed that the key amino acids were Ser330 and Ser387, the main intermolecular forces were hydrogen bond and hydrophobic interaction, and the main binding sites in their molecules were on the beta-lactam ring. Then this protein was combined with streptavidin-labeled tracer and biotinylated fluorescein isothiocyanate to establish a signal-amplified fluorescence polarization assay to determine the 28 drugs in milk. The limits of detection ranged from 0.07 to 2.21 ng/mL, and the sensitivities for the 28 drugs were improved 4 to 48-fold in comparison with the use of a fluorescein isothiocyanate-labeled fluorescent tracer. Therefore, this method could be used for rapid multiscreening of the 28 beta-lactam antibiotics in milk.
引用
收藏
页码:27528 / 27537
页数:10
相关论文
共 38 条
[1]   DETECTION OF β-LACTAM ANTIBIOTICS BASED ON CONJUGATED ANTIBODY WITH GOLD NANORODS BY LOCALIZED SURFACE PLASMON RESONANCE SPECTROMETER [J].
Aghamirzaei, M. ;
Khiabani, M. S. ;
Hamishehkar, H. ;
Mokarram, R. R. ;
Amjadi, M. .
JOURNAL OF APPLIED SPECTROSCOPY, 2022, 89 (02) :391-399
[2]   Anti-Metatype Antibody Screening, Sandwich Immunoassay Development, and Structural Insights for β-Lactams Based on Penicillin Binding Protein [J].
Bai, Yuchen ;
Dou, Leina ;
Wu, Weilin ;
Lu, Zhimin ;
Kou, Jiaqian ;
Shen, Jianzhong ;
Wen, Kai ;
Wang, Zhanhui .
MOLECULES, 2021, 26 (18)
[3]   Design of a sensitive fluorescent polarization immunoassay for rapid screening of milk for cephalexin [J].
Beloglazova, Natalia V. ;
Eremin, Sergei A. .
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2015, 407 (28) :8525-8532
[4]   Epidemiology of β-Lactamase-Producing Pathogens [J].
Bush, Karen ;
Bradford, Patricia A. .
CLINICAL MICROBIOLOGY REVIEWS, 2020, 33 (02)
[5]  
Dabhi M., 2024, J PROTEINSPROTEOMICS, V15, P215, DOI [10.1007/s42485-024-00135-x, DOI 10.1007/S42485-024-00135-X]
[6]   STUDIES ON PLASMA-PROTEIN BINDING OF CARBAMAZEPINE [J].
DISALLE, E ;
PACIFICI, GM ;
MORSELLI, PL .
PHARMACOLOGICAL RESEARCH COMMUNICATIONS, 1974, 6 (02) :193-202
[7]   Recent strategies for inhibiting multidrug-resistant and -lactamase producing bacteria: A review [J].
Fatima, Huma ;
Goel, Nikky ;
Sinha, Rajeshwari ;
Khare, Sunil Kumar .
COLLOIDS AND SURFACES B-BIOINTERFACES, 2021, 205
[8]   Electrochemical Sensor Based on Molecularly Imprinted Polymer for the Detection of Cefalexin [J].
Feier, Bogdan ;
Blidar, Adrian ;
Pusta, Alexandra ;
Carciuc, Paula ;
Cristea, Cecilia .
BIOSENSORS-BASEL, 2019, 9 (01)
[9]   SERINE BETA-LACTAMASES AND PENICILLIN-BINDING PROTEINS [J].
GHUYSEN, JM .
ANNUAL REVIEW OF MICROBIOLOGY, 1991, 45 :37-67
[10]   Direct fluorescence and spectrophotometric-based detection of azithromycin using fluorescein isothiocyanate: Method development and comparative analysis [J].
Hasaneen, Noha ;
Pulicharla, Rama ;
Brar, Satinder Kaur ;
Rezai, Pouya .
JOURNAL OF ENVIRONMENTAL CHEMICAL ENGINEERING, 2024, 12 (03)