Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1168462 | Analytica Chimica Acta | 2009 | 9 Pages |
Abstract
A new phosphorescent labelling reagent consisting of fullerol, fluorescein isothiocyanate and N,N-dimethylaniline (F-ol-(FITC)n-DMA) was developed. The mode of action is based on the reactivity of the active -OH group in F-ol with the -COOH group of FITC to form an F-ol-(FITC)n-DMA complex containing several FITC molecules. F-ol-(FITC)n-DMA increased the number of luminescent molecules in the biological target of WGA-AP-WGA-F-ol-(FITC)n-DMA (WGA and AP are wheat germ agglutinin and alkaline phosphatase, respectively) which improved the sensitivity using solid substrate room temperature phosphorimetry (SSRTP) detection. The proposed method provided high sensitivity and strong specificity for WGA-AP. The limit of detection (LD) was 0.15 ag AP spotâ1 for F-ol and 0.097 ag AP spotâ1 for FITC in F-ol-(FITC)n-DMA, which was lower than the method using single luminescent molecules of F-ol-DMA and FITC-DMA to label WGA (0.20 ag AP spotâ1 for F-ol-DMA and 0.22 ag AP spotâ1 for FITC-DMA). Results for the determination of AP in human serum were in good agreement with those obtained by enzyme-linked immunosorbent assay. The mechanism of F-ol-(FITC)n-DMA labelling of WGA was discussed.
Keywords
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Jia-Ming Liu, Xiao-Mei Huang, Zhen-Bo Liu, Shao-Qin Lin, Fei-Ming Li, Fei Gao, Zhi-Ming Li, Li-Qing Zeng, Lian-Ying Li, Ying Ouyang,