用稳定同位素标记质谱法定量 MUC5AC 和 MUC5B。

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2024-01-01 DOI:10.1007/978-1-0716-3670-1_11

Giorgia Radicioni, Mehmet Kesimer

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引用次数: 0

摘要

粘蛋白 MUC5AC 和 MUC5B 是大型糖蛋白，在上皮表面的先天性防御中发挥着重要作用，对它们在生物样本中的定量分析将有助于了解其来源组织/样本的健康状况。然而，由于它们的大小、异质性和高度糖基化，用传统方法（如 ELISA 和 Western 印迹）对它们进行研究和定量十分困难。我们成功地采用了一种稳定同位素标记质谱方法，对粘蛋白大分子进行绝对定量。在此，我们详细介绍了这种精确、灵敏的液相色谱-质谱（LC-MS）方法，该方法可用于定量分析各种复杂生物样本中的 MUC5AC 和 MUC5B。

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Quantitation of MUC5AC and MUC5B by Stable Isotope Labeling Mass Spectrometry.

Mucins MUC5AC and MUC5B are large glycoproteins that play an essential role in the innate defense of epithelial surfaces and their quantitation in biological samples would be informative about the health status of the tissue/samples they are derived from. However, they are difficult to study and quantify with traditional methods such as ELISA and western blot, due to their size, heterogeneity, and high degree of glycosylation. We successfully implemented a stable isotope labeling mass spectrometry approach for absolute quantification of mucin macromolecules. Here, in detail, we describe this accurate and sensitive liquid chromatography and mass spectrometry (LC-MS) method applied for both MUC5AC and MUC5B quantification in diverse and complex biological samples.

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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.