Determination of alkaline phosphatase based on affinity adsorption solid-substrate room temperature phosphorimetry using rhodamine 6G-dibromoluciferin luminescent nanoparticle to label lectin and prediction of diseases.
In the presence of heavy atom perturber LiAc, the silicon dioxide nanoparticle containing rhodamine 6G (R) and dibromoluciferin (D) (R-D-SiO(2)) can emit strong and stable solid-substrate room temperature phosphorescence signal of R (lambda(ex)/lambda(em)=481/648 nm) and D (lambda(ex)/lambda(em)=457/622 nm) on the surface of acetyl cellulose membrane (ACM). R-D-SiO(2) is used to label triticum vulgare lectin (WGA). Then two types of affinity adsorption reactions, R-D-SiO(2)-WGA- alkaline phosphatase (ALP) (direct method) and WGA-ALP-WGA-R-D-SiO(2) (sandwich method), are carried out on ACM. The conditions and the analytical characteristics for the determination of ALP using affinity adsorption solid-substrate room temperature phosphorimetry (AA-SS-RTP) were studied. For a 0.40-microl drop of sample, results show that the detection limits of the sandwich method are 0.16 ag spot(-1)(457/622 nm) and 0.17 ag spot(-1)(481/648 nm), and the detection limits of the direct method are 0.41 ag spot(-1) (457/622 nm) and 0.44 ag spot(-1) (481/648 nm). The contents of ALP in human serum correlated well with those obtained by enzyme-linked immunoassay. This study shows that AA-SS-RTP whether by the sandwich method or the direct method, can combine very well the characteristics of both high sensitivity of SS-RTP and specificity of the immunoreaction. Simultaneously, whether the phosphorescence excitation/emission wavelength of either R or D in R-D-SiO(2) is chosen to determine ALP, this can promote the agility and widen the adaptability of AA-SS-RTP.
Liu
Jia-Ming
JM
Department of Chemistry and Environment Science, Zhangzhou Normal University, Zhangzhou 363000, PR China.
Liu
Zhen-Bo
ZB
Hu
Li-Xiang
LX
He
Hang-Xia
HX
Yang
Min-Lan
ML
Zhou
Ping
P
Chen
Xin-Hua
XH
Zheng
Min-Min
MM
Zeng
Xiao-Yi
XY
Xu
Yue-Long
YL
D000327:Adsorption; D000469:Alkaline Phosphatase / Q000032:analysis* / Q000378:metabolism; D002846:Chromatography, Affinity; D003955:Diagnostic Tests, Routine / Q000379:methods*; D006801:Humans; D008163:Luminescent Measurements; D053758:Nanoparticles / Q000737:chemistry*; D010756:Phosphoric Acids / Q000032:analysis; D012235:Rhodamines / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*; D013696:Temperature; D014909:Wheat Germ Agglutinins / Q000737:chemistry*
D010756:Phosphoric Acids; D012235:Rhodamines; D014909:Wheat Germ Agglutinins; C026188:rhodamine 6G; D012822:Silicon Dioxide; D000469:Alkaline Phosphatase