Development of a sensitive and accurate enzyme-linked immunosorbent assay (ELISA) system that can replace HPLC analysis for the determination of N1,N12-diacetylspermine in human urine
Development of a sensitive and accurate enzyme-linked immunosorbent assay (ELISA) system that can replace HPLC analysis for the determination of N1,N12-diacetylspermine in human urine
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开发一种灵敏、准确的酶联免疫吸附测定（ELISA）系统，用于替代高效液相色谱法测定人体尿液中的 N1,N12-二乙酰精胺
DOI：
10.1093/oxfordjournals.jbchem.a022086
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发表时间：
1998-07-01
影响因子：
2.7
通讯作者：
Kawakita, M
中科院分区：
文献类型：
--
作者：
Hiramatsu, K;Miura, H;Kawakita, M
N-1,N-12-Diacetylspermine (DiAcSpm)-specific antibodies were raised in rabbits, using N-acetylspermine coupled to mercaptosuccinylated BSA via N-(4-maleimidobutyryloxy)succinimide as an antigen. Highly DiAcSpm-specific antibodies were enriched from crude sera through a series of affinity-based fractionations, A competitive ELISA system, intended for measuring DiAcSpm in solution, was constructed using this antibody preparation, with N-acetylspermine coupled to a synthetic peptide via N-(8-maleimidocapryloxy)succinimide as a solid phase antigen, The K-i value for DiAcSpm with this competitive ELISA system was 33 nM, and the cross-reactivity with DiAcSpm, AcSpm, DiAcSpd, N-1-AcSpd, and N-8-AcSpd was 100, 0.29, 0.20, 0.033, and 0.055%, respectively. This procedure can be applied to the determination of DiAcSpm in human urine samples, giving highly reproducible results. The coefficients of variation obtained were 6.7 and 4.2% for within-run and between-run precision, respectively. The correlation coefficient between DiAcSpm concentrations in urine estimated by ELISA and those by HPLC analysis was calculated to be 0.99, and the regression equation was expressed as y = 1.04x + 0.026 mu M.
