A novel sustainable immunoassay for sensitive detection of atrazine based on the anti-idiotypic nanobody and recombinant full-length antibody

Immunoassays have been widely used to determine small-molecule compounds in food and the environment, meeting the challenge of obtaining false positive or negative results because of the variance in the batches of antibodies and antigens. To resolve this problem, atrazine (ATR) was used as a target,...

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Veröffentlicht in:Chemical engineering journal (Lausanne, Switzerland : 1996). - 1999. - 491(2024) vom: 01. Juli
1. Verfasser: Zhao, Jing (VerfasserIn)
Weitere Verfasser: Li, Peipei, Abd El-Aty, A M, Xu, Lingyuan, Lei, Xingmei, Gao, Song, Li, Jia, Zhao, Yun, She, Yongxin, Jin, Fen, Wang, Jing, Hammock, Bruce D, Jin, Maojun
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2024
Zugriff auf das übergeordnete Werk:Chemical engineering journal (Lausanne, Switzerland : 1996)
Schlagworte:Journal Article Anti-idiotypic nanobody Atrazine Recombinant full-length antibody Specific molecular mechanism Sustainable immunoassay
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100 1 |a Zhao, Jing  |e verfasserin  |4 aut 
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520 |a Immunoassays have been widely used to determine small-molecule compounds in food and the environment, meeting the challenge of obtaining false positive or negative results because of the variance in the batches of antibodies and antigens. To resolve this problem, atrazine (ATR) was used as a target, and anti-idiotypic nanobodies for ATR (AI-Nbs) and a recombinant full-length antibody against ATR (ATR-rAb) were prepared for the development of a sustainable enzyme-linked immunosorbent assay (ELISA). AI-Nb-7, AI-Nb-58, and AI-Nb-66 were selected from an immune phage display library. ATR-rAb was produced in mammalian HEK293 (F) cells. Among the four detection methods explored, the assay using AI-Nb-66 as a coating antigen and ATR-rAb as a detection reagent yielded a half maximal inhibitory concentration (IC50) of 1.66 ng mL-1 for ATR and a linear range of 0.35-8.73 ng mL-1. The cross-reactivity of the assay to ametryn was 64.24%, whereas that to terbutylazine was 38.20%. Surface plasmon resonance (SPR) analysis illustrated that these cross-reactive triazine compounds can bind to ATR-rAb to varying degrees at high concentrations; however, the binding/dissociation kinetic curves and the response values at the same concentration are different, which results in differences in cross-reactivity. Homology modeling and molecular docking revealed that the triazine ring is vital in recognizing triazine compounds. The proposed immunoassay exhibited acceptable recoveries of 84.40-105.36% for detecting fruit, vegetables, and black tea. In conclusion, this study highlights a new strategy for developing sustainable immunoassays for detecting trace pesticide contaminants 
650 4 |a Journal Article 
650 4 |a Anti-idiotypic nanobody 
650 4 |a Atrazine 
650 4 |a Recombinant full-length antibody 
650 4 |a Specific molecular mechanism 
650 4 |a Sustainable immunoassay 
700 1 |a Li, Peipei  |e verfasserin  |4 aut 
700 1 |a Abd El-Aty, A M  |e verfasserin  |4 aut 
700 1 |a Xu, Lingyuan  |e verfasserin  |4 aut 
700 1 |a Lei, Xingmei  |e verfasserin  |4 aut 
700 1 |a Gao, Song  |e verfasserin  |4 aut 
700 1 |a Li, Jia  |e verfasserin  |4 aut 
700 1 |a Zhao, Yun  |e verfasserin  |4 aut 
700 1 |a She, Yongxin  |e verfasserin  |4 aut 
700 1 |a Jin, Fen  |e verfasserin  |4 aut 
700 1 |a Wang, Jing  |e verfasserin  |4 aut 
700 1 |a Hammock, Bruce D  |e verfasserin  |4 aut 
700 1 |a Jin, Maojun  |e verfasserin  |4 aut 
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