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首页> 外文期刊>Analytical Biochemistry: An International Journal of Analytical and Preparative Methods >Development and validation of analytical methodologies for the quantification of PCK3145 and PEG-PCK3145 in mice
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Development and validation of analytical methodologies for the quantification of PCK3145 and PEG-PCK3145 in mice

机译：小鼠PCK3145和PEG-PCK3145定量分析方法的开发与验证

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摘要

PCK3145 is an anti-metastatic synthetic peptide against prostate cancer. The objective of the study is to develop and validate novel and sensitive methods for the determination of PCK3145 and Pegylated PCK3145 in mouse plasma. An LC–MS/MS method was developed and validated for the determination of PCK3145 giving high sensitivity and linearity in the range of 0.125–4.0 μg/mL. PCK3145 characterised by short half-life, therefore, it was conjugated with the poly ethylene glycol (PEG). However, LC-MS/MS has been more difficult to apply for the quantitative analysis of PEGylated peptides due to the large size. A UHPLC-UV method was developed and validated for the determination of PEG-PCK3145, with linearity of 0.05–2.0?mg/mL. In order to further improve the sensitivity for the detection of PEG-PCK3145, an indirect ELISA method was used. It was found that this method was capable of detecting PCK3145 through the quantification of PEG with excellent sensitivity found at 0.132?ng/mL. Thein vitroproteolytic stability of PCK3145 and PEG-PCK3145 in mouse plasma and whole blood was studied by LC-MS/MS and UHPLC, respectively. The LC–MS/MS and ELISA methods can be applied for monitoring levels of PCK3145 in mouse plasma forin vivopharmacokinetic and bioavailability animal studies.
机译：PCK3145是一种针对前列腺癌的抗转移性合成肽。该研究的目的是开发和验证用于测定小鼠等离子体中PCK3145和PEG化PCK3145的新颖和敏感方法。开发并验证了LC-MS / MS方法，用于测定PCK3145，在0.125-4.0μg/ ml的范围内提供高灵敏度和线性度。因此，PCK3145特征在于半寿命，因此，与聚乙二醇（PEG）缀合。然而，由于大尺寸，LC-MS / MS更难以施加对聚乙二醇化肽的定量分析。开发UHPLC-UV方法并验证用于测定PEG-PCK3145，线性度为0.05-2.0×mg / ml。为了进一步提高PEG-PCK3145检测的灵敏度，使用间接ELISA方法。发现该方法能够通过PEG的定量检测PCK3145，优异的敏感性在0.132Ω·Ng / mL。通过LC-MS / MS和UHPLC研究了小鼠血浆和全血中PCK3145和PEG-PCK3145的培养素衍生稳定性。 LC-MS / MS和ELISA方法可用于监测小鼠血浆肢体动力学和生物利用性动物研究中PCK3145的水平。

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