[Establishment of an ELISA system of N1,N12-diacetylspermine in human urine].
OBJECTIVE
It is known that two kinds of Diacetylpolyamine, N1,N12-Diacetylspermine(DiAcSpm) and N1,N8-Diacetylspermidine(DiAcSpd), are excreted in urine. Although these two substances are 0.4% and 1.2% of the whole polyamine, respectively, these substances may be important for a sick diagnosis. The aim of this study was to develop a sensitive and specific method for detecting DiAcSpm.


METHODS
We developed a new competitive ELISA system for the measurement of DiAcSpm in human urine, using polyclonal antibodies against DiAcSpm.


RESULTS
The lower limit of detection of this ELISA was 4.53 nM/assay. The higher limit of detection was 145 nM/assay. Mean recovery was 101% (range, 96.3-108%). The coefficients of variation (CV) for within-run measurements by this method were 4.87% (mean = 95.5 nM) and 5.20% (mean = 32.4 nM), and the between-run CV were 7.53% (mean = 101 nM) and 9.46% (mean = 33.8 nM).


CONCLUSIONS
We have established an ELISA system for the quantification of urinary DiAcSpm that uses novel polyclonal antibodies. Our ELISA system is simple and sensitive.
Matsuoka
Akemi
A
Trans Genic Inc., Kamimashiki-pref., Kumamoto 861-2202.
Sakamoto
Tamami
T
D014408:Biomarkers, Tumor / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D011933:Reagent Kits, Diagnostic / Q000592:standards*; D012680:Sensitivity and Specificity; D013095:Spermidine / Q000031:analogs & derivatives* / Q000652:urine*; D013096:Spermine / Q000031:analogs & derivatives* / Q000652:urine*
D014408:Biomarkers, Tumor; C093909:N(1),N(8)-diacetylspermidine; D011933:Reagent Kits, Diagnostic; D013096:Spermine; C029453:N',N''-diacetylspermine; D013095:Spermidine