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Ultrasensitive detection of ferritin in human serum by Western blotting based on quantum dots-labeled avidin-biotin system
Authors:Liu Pingping  Na Na  Liu Tingting  Huang Lingyun  He Dacheng  Hua Wenhao  Ouyang Jin
Affiliation:College of Chemistry, Beijing Normal University, Beijing, P R China.
Abstract:
Various methods have been applied for serum ferritin detection, however, these methods still have some limitations. Over the last few years, quantum dots (QDs) have become very attractive for immunoassays because of their enormous potentials in ultrasensitive analysis. In this study, a Western blotting method combined with QDs‐labeled avidin–biotin system for detecting human serum ferritin was described. Meanwhile, the traditional diaminobenzidine (DAB)–horseradish peroxidase (HRP) method had been compared with the method. The linearity of this QDs‐based Western blotting method was from 0.27 to 1.1 ng, and the quantification limit was 0.27 ng, the sensitivity was up to pictogram values. Real serum samples such as hepatoma, thalassemia patient and normal individual sera were analyzed, the analysis results demonstrated that there was significant difference in the concentrations of ferritin between patients and normal individual serum. Furthermore, the recovery of ferritin from the serum samples of patients ranged from 98.15 to 119.67%, and the RSD (relative standard deviation) ranged from 8.73 to 11.61%, the repeatabilities were well within the acceptable range, which revealed that this method is a stable and reproducible method for detecting serum ferritin and have potential application prospect in clinical laboratory of serum ferritin detection.
Keywords:Biomedicine  Ferritin  Hepatoma  Quantum dots  Thalassemia  Western blotting
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