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Development of a Colloidal Gold-based Immunochromatographic Test Strip for Detection of Cetacean Myoglobin
Published on: July 13, 2016
A highly sensitive and specific immunochromatographic test strip for detecting p,p'-DDA in seawater and shellfish
Teng Zhang1, Yunrui Xing2, Jiale Yao1
1Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, School of Life Science and Agricultural Engineering, Nanyang Normal University, Nanyang 473000, China.
None:
P,p'-DDA, similar to DDT, accumulates in the food chain and poses potential risks to human health. Herein, a novel sensitive and specific mAb-10B4D7 was screened and subsequently rAb-10B4D7 was expressed by genetic engineering. Utilizing rAb-10B4D7, an immunochromatographic test strip (ITS) was developed with visual limit of detection 12.5 μg/L within 15 min, displaying no cross-reactivity with o,p'-DDT, p,p'-DDT, p,p'-DDD, and p,p'-DDE. Further, a linear relationship between log(concentration of p,p'-DDA) and the optical density ratio of test (T) line and control (C) line was established with limits of detection of 1.03 μg/kg, recoveries of 79.3-106.8 %, and coefficients of variation below 10 %. Molecular docking simulations identified two critical residues (Arg-H62, Phe-L98) forming three hydrogen bonds with p,p'-DDA, resulting in the high specificity and affinity of rAb-10B4D7, with a KD of 5.81 × 10-9 mol/L. These results indicated that the ITS was suitable for monitoring p,p'-DDA and organochlorine residues in seawater and shellfish.
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