Construction of Immunomagnetic Particles with High Stability in Stringent Conditions by Site-Directed Immobilization of Multivalent Nanobodies onto Bacterial Magnetic Particles for the Environmental Detection of Tetrabromobisphenol-A
2019
He, Jinxin | Ma, Shijiao | Wu, Sha | Xu, Junjie | Tian, Jiesheng | Li, Ji | Gee, Shirley J. | Hammock, Bruce D. | Li, Qing X. | Xu, Ting
Bacterial magnetic particles (BMPs) are an attractive carrier material for immunoassays because of their nanoscale size, dispersal ability, and membrane-bound structure. Antitetrabromobisphenol-A (TBBPA) nanobodies (Nbs) in the form of monovalence (Nb1), bivalence (Nb2), and trivalence (Nb3) were biotinylated and immobilized onto streptavidin (SA)-derivatized BMPs to construct the complexes of BMP-SA-Biotin-Nb1, -Nb2, and -Nb3, respectively. An increasing order of binding capability of BMP-SA-Biotin-Nb1, -Nb2, and -Nb3 to TBBPA was observed. These complexes showed high resilience to temperature (90 °C), methanol (100%), high pH (12), and strong ionic strength (1.37 M NaCl). A BMP-SA-Biotin-Nb3-based enzyme linked immunosorbent assay (ELISA) for TBBPA dissolved in methanol was developed, showing a half-maximum inhibition concentration (IC₅₀) of 0.42 ng mL–¹. TBBPA residues in landfill leachate, sewage, and sludge samples determined by this assay were in a range of <LOD−1.17 ng mL–¹, <LOD−0.75 ng mL–¹, and <LOD−3.65 ng g–¹ (dw), respectively, correlating well with the results by liquid chromatography tandem mass spectrometry. The BMP-SA-Biotin-Nb3 was reusable at least three times without significant loss of the binding capability. The BMP-SA-Biotin-Nb3-based ELISA, with a total assay time of less than 30 min, is promising for the rapid monitoring of TBBPA in the environment.
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