The kidney is one of the major target organs for drug-induced toxicity. During drug development, the traditional markers of nephrotoxicity indicate only severe and late damage, which leads to high costs. The new biomarkers are needed for a more sensitive and reliable evaluation of nephrotoxicity, especially for the regulatory accepted and validated in vitro model. We developed an in vitro model based on the HK-2 cell using the biomarkers of nephrotoxicity as endpoints for the evaluation of nephrotoxicity. The predictive performance of the biomarkers including LDH, GGT, KIM-1, clusterin, CysC, NGAL, TIMP-1, GSTπ and osteopontin was evaluated with 22 well characterized compounds. The area under the curve (AUC) values of KIM-1, clusterin, CysC and osteopontin ranged between 0.79 and 0.84. The combination of clusterin, KIM-1 and/or osteopontin improved the AUC value (ranging between 0.88 and 0.95) compared to one biomarker. Taken together, these results suggest that the model based on the HK-2 cell using clusterin, osteopontin, CysC and KIM-1 as endpoints would allow the prediction of nephrotoxicity at early preclinical stages.
Jing Wang,Kai Mu,Hongjuan Wei,Hong Chen,Yunxiang Wang,Wuxing Zhang,Zhen Rong
Acute kidney injury (AKI) is a serious renal emergency that is often underdiagnosed and associated with high mortality rates. Early diagnosis of AKI is crucial to improve patient prognosis and reduce healthcare costs. In this study, a paper-based colloidal gold multiplex vertical flow assay was developed for rapid detection of three AKI biomarkers, namely, NGAL , CysC, and RBP4 , in a single test kit. In this system, specific capture antibodies were pre-immobilized on the sensing membrane at four spatially isolated immunoreactive sites. Gold nanoparticles were conjugated with detection antibodies as the colorimetrical labels. The resultant images could be read out qualitatively by the naked eyes or quantitatively by a smartphone-based readout device. Three AKI biomarkers could be simultaneously detected in 10–15 min. The detection sensitivity and specificity for clinical serum samples collected from 25 patients with AKI and 25 healthy individuals were determined as 92% and 88%. This vertical flow assay platform is easy to use, rapid, and low cost. It is a promising diagnostic tool for multiplex detection of disease biomarkers at point of care.
Nanfei Yang,Yuncheng Bei,Yahong Huang,Wei Zheng,Jiehua Ma,Jiangqiong Ke
Serum Cystatin C (CysC) is an impressive marker for early diagnosis of renal dysfunction. In this work, we established a novel electrochemical immunosensor based on Fe 3 O 4 /AuNPs-MWCNTs@PDA nanocomposite for the detection of CysC. The Fe 3 O 4 /AuNPs-MWCNTs@PDA nanozyme complex by polydopamine encapsulation can not only carry massive detection antibodies, but also bind the electroactive substance toluidine blue (TB) through electrostatic adsorption. By immobilizing AuNPs onto the electrode to bind the capture antibody (Ab1), we constructed a sandwich electrochemical immunosensor with low cost, high sensitivity, and repeatability. The detection range is 3.9–125.0 ng/mL with a significant linear relationship between the current peak difference (ip) and logarithm of the CysC concentration. Moreover, the detection limit of the immunosensor is 0.157 ng/mL. We have successfully utilized this novel immunosensor to detect CysC in human serum samples, and these results have implications for its potential use in clinical application.
危险品化学品经营许可证（带存储）