Determination of trace alkaline phosphatase by solid-substrate room-temperature phosphorimetry based on Triticum vulgare lectin labeled with fullerenol.
Fullerenol (F) shows a strong and stable room-temperature phosphorescence (RTP) signal on the surface of nitrocellulose membrane (NCM) at lambda ex max/ lambda em max =542.0/709.4 nm. When modified by dodecylbenzenesulfonic acid sodium salt (DBS), fullerenol emits a stronger signal. It was also found that quantitative specific affinity-adsorption reaction can be carried out between Triticum vulgare lectin (WGA) labeled with DBS-F and alkaline phosphatase (ALP) on the surface of NCM, and the product obtained (WGA-ALP-WGA-F-DBS) emits a strong and stable RTP signal. Furthermore, the content of ALP was proportional to the DeltaI(p) value. Based on the facts above, a new method for the determination of trace amounts of ALP by affinity-adsorption solid-substrate room-temperature phosphorimetry (AA-SS-RTP) was established, using fullerenol modified with DBS to label WGA. The detection limit was 0.011 fg spot(-1) (corresponding concentration: 2.8x10(-14) g ml(-1), namely 2.8x10(-16) mol l(-1)). This method with high sensitivity, accuracy, and precision has been successfully applied to the determination of the content of ALP in human serum survey and forecast human disease, and the results are tallied with those using alkaline phosphatase kits. The mechanism for the determination of ALP using AA-SS-RTP was also discussed.
Liu
Jia-Ming
JM
Department of Chemistry and Environment Science, Zhangzhou Normal University, Zhangzhou 363000, P. R. China.
Gao
Fei
F
Huang
Hong-Hua
HH
Zeng
Li-Qing
LQ
Huang
Xiao-Mei
XM
Zhu
Guo-Hui
GH
Li
Zhi-Ming
ZM
D000328:Adult; D000469:Alkaline Phosphatase / Q000032:analysis*; D001557:Benzenesulfonates; D003101:Collodion; D005260:Female; D037741:Fullerenes / Q000737:chemistry*; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D008567:Membranes, Artificial; D014909:Wheat Germ Agglutinins / Q000737:chemistry*
D001557:Benzenesulfonates; D037741:Fullerenes; D008567:Membranes, Artificial; D014909:Wheat Germ Agglutinins; C108127:fullerenol; C001114:dodecylbenzenesulfonic acid; D003101:Collodion; D000469:Alkaline Phosphatase