Enhancing the detection sensitivity of nanobody against aflatoxin B1 through structure-guided modification

被引:18
作者
He, Ting [1 ]
Nie, Yao [1 ,2 ]
Yan, Tingting [1 ,2 ]
Zhu, Jiang [1 ,2 ]
He, Xiaoling [1 ]
Li, Ying [1 ,2 ]
Zhang, Qi [3 ]
Tang, Xiaoqian [3 ]
Hu, Rui [1 ,2 ]
Yang, Yunhuang [1 ,2 ]
Liu, Maili [1 ,2 ]
机构
[1] Huazhong Univ Sci & Technol, Hubei Opt Valley Lab,Key Lab Magnet Resonance Bio, Chinese Acad Sci,Wuhan Natl Lab Optoelect,State K, Wuhan Inst Phys & Math,Innovat Acad Precis Measur, Wuhan 430071, Peoples R China
[2] Univ Chinese Acad Sci, Beijing 100049, Peoples R China
[3] Chinese Acad Agr Sci, Oil Crops Res Inst, Wuhan 430062, Peoples R China
基金
中国国家自然科学基金;
关键词
Nanobody; Aflatoxin B-1; Structure-guided modification; SINGLE-DOMAIN ANTIBODY; AFFINITY IMPROVEMENT; PROTEIN STRUCTURES; BINDING-SITE; VHH; IMMUNOASSAY; RESIDUES; DYNAMICS; SEQUENCE; SYSTEM;
D O I
10.1016/j.ijbiomac.2021.11.182
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Nanobodies (Nbs) have shown great potential in immunodetection of small-molecule contaminants in food and environmental monitoring. However, the limited knowledge of the mechanism of Nbs binding to small molecules has hampered the development of high-affinity Nbs and assay improvement. We previously reported two homologous nanobodies Nb26 and Nb28 specific to aflatoxin B-1 (AFB(1)), with the former exhibiting higher sensitivity in ELISA. Herein, Nb26 was selected as the model antibody to resolve its solution nuclear magnetic resonance (NMR) structure, and investigate its AFB(1) recognition mechanism. The results revealed that Nb26 exhibits a typical immunoglobulin fold and its AFB(1)-binding interface is uniquely located in complementarity-determining region 3 (CDR3) and framework region 2 (FR2). This finding was applied to improve the binding activity of Nb28 against AFB(1) by constructing two Nb28-based mutants A(50)V and (SD)-D-102, resulting in 2.3- and 3.3-fold sensitivity enhancement over the wild type, respectively. This study develops an NMR-based strategy to analyze the underlying mechanism of Nb against AFB(1), and successfully generated two site-modified Nbs with improved detection sensitivity. It is believed that this work could greatly expand the applications of Nbs by providing a way to enhance the binding activity.
引用
收藏
页码:188 / 197
页数:10
相关论文
共 50 条
[41]   An aptamer-based dipstick assay for the rapid and simple detection of aflatoxin B1 [J].
Shim, Won-Bo ;
Kim, Min Jin ;
Mun, Hyoyoung ;
Kim, Min-Gon .
BIOSENSORS & BIOELECTRONICS, 2014, 62 :288-294
[42]   GO-amplified fluorescence polarization assay for high-sensitivity detection of aflatoxin B1 with low dosage aptamer probe [J].
Ye, Hua ;
Lu, Qianqian ;
Duan, Nuo ;
Wang, Zhouping .
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2019, 411 (05) :1107-1115
[43]   Assessment of the Ameliorative Effect of Bacillus subtilis against the Toxicity Induced by Aflatoxin B1 in Rats [J].
Deabes, Mohamed M. ;
Aboulthana, Wael M. ;
Ahmed, Kawkab A. ;
Naguib, Khayria M. .
EGYPTIAN JOURNAL OF CHEMISTRY, 2021, 64 (04) :2141-2164
[44]   The effect of different stabilizers on stability of horseradish peroxidase-bovine serum albumin-aflatoxin B1, a conjugated tracer for detection of aflatoxin B1 in immunoassay-based methods [J].
Farzamfar, Bardia ;
Bayanolhagh, Saeed ;
Mahboudi, Fereidoun ;
Zahrai, Mahin .
IRANIAN JOURNAL OF PHARMACEUTICAL RESEARCH, 2007, 6 (03) :179-184
[45]   Using of Tyramine Signal Amplification to Improve the Sensitivity of ELISA for Aflatoxin B1 in Edible Oil Samples [J].
Zhang, Yuanyuan ;
Li, Ming ;
Cui, Yin ;
Hong, Xia ;
Du, Daolin .
FOOD ANALYTICAL METHODS, 2018, 11 (09) :2553-2560
[46]   Accelerated Hybridization Chain Reaction Kinetics Using Poly DNA Tetrahedrons and Its Application in Detection of Aflatoxin B1 [J].
Li, Xuesong ;
Cheng, Jie ;
Zeng, Kun ;
Wei, Shulin ;
Xiao, Jiaxuan ;
Lu, Yanyan ;
Zhu, Fang ;
Wang, Zhanhui ;
Wang, Kun ;
Wu, Xiangyang ;
Zhang, Zhen .
ACS APPLIED MATERIALS & INTERFACES, 2023, 15 (35) :41237-41246
[47]   An Antibody Ordered Assembly Functional BPE-ECL Platform for Aflatoxin B1 Detection [J].
Qiao, Fanglin ;
Chen, Yin ;
Fei, Yunjie ;
Jiang, Yanjun ;
Xiong, Xiaohui ;
Liu, Yuanjian .
FOOD ANALYTICAL METHODS, 2024, 17 (02) :260-268
[48]   Sensitive Aflatoxin B1 Detection Using Nanoparticle-Based Competitive Magnetic Immunodetection [J].
Pietschmann, Jan ;
Spiegel, Holger ;
Krause, Hans-Joachim ;
Schillberg, Stefan ;
Schroeper, Florian .
TOXINS, 2020, 12 (05)
[49]   Development of a Lateral Flow Strip with a Positive Readout for the On-Site Detection of Aflatoxin B1 [J].
Shen, Kemin ;
Hu, Xiaoqin ;
Sun, Linlin ;
Han, Chun ;
Yang, Jianzhou .
MOLECULES, 2022, 27 (15)
[50]   Electrochemical detection of aflatoxin B1: an aptasensor prepared using graphene oxide and gold nanowires [J].
Nodoushan, Somayeh Mousavi ;
Nasirizadeh, Navid ;
Kachuei, Reza ;
Fooladi, Abbas Ali Imani .
ANALYTICAL METHODS, 2019, 11 (47) :6033-6042