用于定量分析自发性痰液中MUC5AC和MUC5B粘蛋白的液相色谱-串联质谱法的开发与验证
Development and qualification of an LC-MS/MS method for quantification of MUC5AC and MUC5B mucins in spontaneous sputum.
作者信息
Sun Weiwen, Mou Si, Huntington Catherine, Killick Helen, Scott Ian Christopher, Kelly Aoife, Gavala Monica, Larsson Jessica, Vakkalanka Mani Deepika, Alexis Neil E, Wiley Walter, Wheeler Aaron, Shah Kumar, Yuan Moucun, Mylott William R, Contrepois Kévin, Rosenbaum Anton I
机构信息
Integrated Bioanalysis, Clinical Pharmacology and Safety Sciences, R&D, AstraZeneca, South San Francisco, CA, USA.
Biologics Engineering, R&D, AstraZeneca, Cambridge, USA.
出版信息
Bioanalysis. 2025 Feb;17(3):187-198. doi: 10.1080/17576180.2025.2457844. Epub 2025 Feb 20.
AIM
Airway mucins in sputum are promising respiratory disease biomarkers, despite posing substantial analytical challenges due to their physicochemical properties and rare and heterogenous nature of the matrix. We aimed to identify a suitable sputum collection and processing method, and qualify a bioanalytical method for MUC5AC and MUC5B quantification in clinical samples.
METHOD
Mucins were quantified in induced and spontaneous sputum collected from the same COPD patients, following various sample processing procedures. LC-MS/MS method used truncated recombinant mucins as surrogate analytes in surrogate matrix.
RESULTS
Frozen spontaneous sputum was found to be a suitable and convenient matrix for mucin quantification and fit-for-purpose method qualification was performed.
CONCLUSION
Our methodology provides accurate and reliable MUC5AC and MUC5B quantification and facilitates multi-site clinical sputum collection.
目的
痰液中的气道黏蛋白是很有前景的呼吸系统疾病生物标志物，尽管由于其物理化学性质以及基质的稀有和异质性给分析带来了巨大挑战。我们旨在确定一种合适的痰液采集和处理方法，并验证一种用于临床样本中MUC5AC和MUC5B定量分析的生物分析方法。
方法
对同一慢性阻塞性肺疾病（COPD）患者诱导痰和自然咳出痰采用各种样本处理程序后进行黏蛋白定量分析。液相色谱-串联质谱（LC-MS/MS）方法使用截短的重组黏蛋白作为替代基质中的替代分析物。
结果
发现冷冻的自然咳出痰是黏蛋白定量分析合适且方便的基质，并进行了适用方法验证。
结论
我们的方法可实现准确可靠的MUC5AC和MUC5B定量分析，并有助于多中心临床痰液采集。
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