重组红细胞生成素检测内部标准的研究。

IF 2.6 3区 医学 Q2 BIOCHEMICAL RESEARCH METHODS
Chengshuai Niu, Kaifeng Liu, Xinchao Liu, Sen He, Xinmiao Zhou
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引用次数: 0

摘要

随着EPO基因c.577del变异的发现,检测重组红细胞生成素(rEPO)的过程变得复杂而耗时。为了解决这一问题,我们建立了一种利用反正常免疫纯化联合western blotting (WB)和n -月桂酰肌氨酸钠聚丙烯酰胺凝胶电泳(SAR-PAGE)检测EPO变体(VAR-EPO)和具有抗VAR-EPO和抗EPO抗体的rEPO的rEPO确认方法。因此,有必要开发一种能够被抗var - epo抗体识别的内标(is)来监测反向免疫纯化,确保分析的可靠性和准确性。在本研究中,我们构建了一个基于聚乙二醇修饰VAR-EPO的IS,并评估了其在兴奋剂分析中的可靠性和适用性。本文提供的数据表明,所获得的聚乙二醇化VAR-EPO不仅可以作为IS监测尿液样本中rEPO和VAR-EPO的检测,还可以监测EPO受体激动剂的检测。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Study on Internal Standards Applicable to the Detection of Recombinant Erythropoietin.

With the discovery of the variant c.577del in the EPO gene, the procedure for detecting the presence of recombinant erythropoietin (rEPO) has become complicated and time-consuming. To address this situation, an rEPO confirmation method that uses reverse-normal immunopurification coupled with western blotting (WB) and sodium N-lauroylsarcosinate polyacrylamide gel electrophoresis (SAR-PAGE), which can detect the EPO variant (VAR-EPO) and rEPO with anti-VAR-EPO and anti-EPO antibodies, has been developed. Therefore, it is necessary to develop an internal standard (IS) that can be recognized by an anti-VAR-EPO antibody to monitor reverse immunopurification, ensuring the reliability and accuracy of the analysis. In this study, we constructed an IS based on VAR-EPO modified with polyethylene glycol (PEG) and then assessed its reliability and applicability in doping analysis. The data provided here show that the obtained PEGylated VAR-EPO can be used as an IS to monitor the detection of not only rEPO and VAR-EPO but also EPO receptor agonists in urine samples.

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来源期刊
Drug Testing and Analysis
Drug Testing and Analysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL
CiteScore
5.90
自引率
24.10%
发文量
191
审稿时长
2.3 months
期刊介绍: As the incidence of drugs escalates in 21st century living, their detection and analysis have become increasingly important. Sport, the workplace, crime investigation, homeland security, the pharmaceutical industry and the environment are just some of the high profile arenas in which analytical testing has provided an important investigative tool for uncovering the presence of extraneous substances. In addition to the usual publishing fare of primary research articles, case reports and letters, Drug Testing and Analysis offers a unique combination of; ‘How to’ material such as ‘Tutorials’ and ‘Reviews’, Speculative pieces (‘Commentaries’ and ‘Perspectives'', providing a broader scientific and social context to the aspects of analytical testing), ‘Annual banned substance reviews’ (delivering a critical evaluation of the methods used in the characterization of established and newly outlawed compounds). Rather than focus on the application of a single technique, Drug Testing and Analysis employs a unique multidisciplinary approach to the field of controversial compound determination. Papers discussing chromatography, mass spectrometry, immunological approaches, 1D/2D gel electrophoresis, to name just a few select methods, are welcomed where their application is related to any of the six key topics listed below.
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