一种快速检测人血清中尿酸酶活性方法的建立与验证
Establishment and Validation of a Rapid Method for Detecting Uricase Activity in Serum of Human
霍一凡 1满素勤 2徐嘉辰 2徐婷婷 1邱云良2
作者信息
- 1. 中国医药工业研究总院,上海 201203;上海益诺思生物技术股份有限公司,上海 201203
- 2. 上海益诺思生物技术股份有限公司,上海 201203
- 折叠
摘要
基于尿酸酶催化尿酸分解产生尿囊素、过氧化氢和二氧化碳的原理,该研究建立了在37 ℃、pH 7.5的反应体系中,检测尿酸酶活性的方法.方法学验证结果表明,该方法对过氧化氢和尿酸酶的定量范围分别为0.049～100.000 μmol/L、0.819～200.000mU/mL;方法的批内、批间精密度的变异系数小于20.0％,准确度相对误差在±25.0％范围内;高血脂(血脂指数3 mg/mL)样品和2％溶血个体样品相对于普通个体的回收率结果显示,高血脂对尿酸酶活性检测无干扰,而溶血有干扰;人血清尿酸酶样品于室温或2～8 ℃放置不超过24h、冻融不超过3次、-70～-90 ℃放置不超过64 d,均不影响尿酸酶活性.该方法灵敏度高、精密度好、准确度高,且简便、快速,可支持尿酸酶药物的临床前及临床评价.
Abstract
Based on the principle that uricase catalyzes the decomposition of uric acid to produce allantoin,hydrogen peroxide and carbon dioxide,this paper established a method to detect the activity of uricase at 37 ℃ and pH 7.5.The results showed that the quantitative ranges of hydrogen peroxide and uricase were 0.049-100.000 μmol/L and 0.819-200.000 mU/mL,respectively.The within-and between-run variation coefficients of precision were less than 20.0％,and the relative error of accuracy were within±25.0％.The results of recovery experiments using hyperlipemia samples(lipid index 3 mg/mL)and 2％hemolysis donor samples compared to normal donor samples showed that hyperlipemia did not interfere with the detection of uricase activity,whereas hemolysis did.Uricase activity was not affected when human serum uricase samples were placed at room temperature or 2-8 ℃ for no more than 24 h,frozen and thawing for no more than 3 cycles,and stored at-70-90 ℃ for no more than 64 days.In conclusion,the established method is sensitive,precise,accurate,simple,rapid,and can support the evaluation of preclinical and clinical studies of uricase drugs.
关键词
尿酸酶/尿酸/过氧化氢/酶促反应/尿酸酶活性Key words
uricase/uric acid/hydrogen peroxide/enzymatic reaction/uricase activity引用本文复制引用
出版年
2024
NETL
NSTL
万方数据