Determination of 3-nitrotyrosine in human urine samples by surface plasmon resonance immunoassay |
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Authors: | Jing JinAuthor Vitae Chunyan WangAuthor Vitae Yi TaoAuthor VitaeYingjun TanAuthor Vitae Dacan YangAuthor VitaeYin GuAuthor Vitae Huihua DengAuthor VitaeYanqiang BaiAuthor Vitae Hua LuAuthor Vitae Yumin WanAuthor VitaeZuhong LuAuthor Vitae Yinghui LiAuthor Vitae |
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Affiliation: | a Chien-Shiung Wu Laboratory, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China b Key Laboratory of Child Development and Learning Science (Southeast University), Ministry of Education, Nanjing 210096, China c State Key Laboratory of Space Medicine Fundamentals and Application, Chinese Astronaut Research and Training Center, China |
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Abstract: | This paper describes the detection of a low-molecular weight molecule, 3-nitrotyrosine (3-NT) (∼226 Da), in human urine by coupling indirect inhibition assay with a surface plasmon resonance (SPR) sensor. 3-NT antibody (anti-3-NT Ab, mouse IgG) was used in this assay. An optimal antibody concentration has been measured at 27.9 μg/mL in order to obtain the best performance of the sensor surface. The lowest detection limit for 3-NT with this method is 4.7 ng/mL (S/N = 3). Sensor reliability was demonstrated by good specificity, intra-assay and inter-assay relative standard deviations <8%, average recovery of 107.68 ± 19.4% and sensor surface (self-assembled monolayer) stability through more than 200 regeneration cycles and 15 days of repeated measurement. This is the first SPR biosensor assay of 3-NT in human urine. The high stability of the SPR sensor surface underlies the potential of the SPR method as a low cost diagnostic tool for clinical detection of 3-NT. |
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Keywords: | Surface plasmon resonance Self-assembled monolayer 3-Nitrotyrosine Urine Immunosensor |
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