Advances in LC-MS/MS for Biological Quantitative Analysis of Protein Drugs
液相色谱-串联质谱（LC-MS/MS）在生物蛋白药物定量分析中的进展
摘要 (Abstract)
1. Biomed Pharmacother. 2025 Nov;192:118558. doi: 10.1016/j.biopha.2025.118558. Epub 2025 Sep 18. Development and application of a multimatrix LC-MS/MS method for quantifying elexacaftor-tezacaftor-ivacaftor: Expanding therapeutic drug monitoring in cystic fibrosis from systemic circulation to airways and sweat. Mucci M(1), Colarelli M(2), Ripani P(3), Di Nicola M(2), Del Ciotto M(3), Di Sabatino M(3), Collini F(3), Romano M(2), Recchiuti A(4). Author information: (1)Department of Medical, Oral, and Biotechnology Science (DSMOB), G. d'Annunzio University of Chieti - Pescara, Center of Advanced Studies and Technology, via Polacchi 13, Chieti 66100, Italy. Electronic address: matteo.mucci@unich.it. (2)Department of Medical, Oral, and Biotechnology Science (DSMOB), G. d'Annunzio University of Chieti - Pescara, Center of Advanced Studies and Technology, via Polacchi 13, Chieti 66100, Italy. (3)Regional CF Center, San Liberatore Hospital - ASL Teramo, Atri, Italy. (4)Department of Medical, Oral, and Biotechnology Science (DSMOB), G. d'Annunzio University of Chieti - Pescara, Center of Advanced Studies and Technology, via Polacchi 13, Chieti 66100, Italy. Electronic address: antonio.recchiuti@unich.it. BACKGROUND: Therapeutic drug monitoring (TDM) is becoming increasingly essential in cystic fibrosis (CF), as a growing number of children and adults are now eligible for elexacaftor/tezacaftor/ivacaftor (ETI) CF Transmembrane conductance Regulator (CFTR) modulator therapies. Although plasma remains the benchmark, its invasive nature limits practical use. Analytical approaches based on dried matrices currently available still rely on venipuncture and intensive workflows. To overcome these limitations, we developed an advanced, multidimensional TDM framework for ETI quantification that incorporates truly venipuncture-free, self-collected, and quantitative dried blood spot (DBS) sampling, along with non-invasive nasal airway swabs (NAS) and sweat. We also evaluated ETI in airway and sweat fluids, offering insight into drug distribution and activity at key target tissues. METHODS: Selectivity, specificity, linearity, accuracy, precision, and inter/intraday stability of the methods applied to plasma, NAS, and sweat specimens were investigated according to ICH M10 guidelines and IATDMCT for DBS. RESULTS: Analytical performance of the methods in all tested matrices were demonstrated and met the acceptance criteria of ICH M10 guidelines for bioanalytical-method validation, with DBS additionally fulfilling the IATDMCT DBS-specific recommendations. DBS measurements were statistically equivalent to plasma concentrations, supporting their utility as a minimally invasive surrogate for TDM. ETI levels in NAS were significantly higher than in plasma or sweat, indicating localized accumulation of the drug on the airway surface, a key site of CFTR activity. Sweat samples, while showing lower analyte amounts, contained detectable levels in all analytes within their biologically active concentrations. CONCLUSIONS: This integrated analytical approach provides a holistic view of systemic and local ETI distribution, increasing the potential for personalized TDM, pharmacokinetics/pharmacodynamics study, and optimization of CFTR modulator therapy. Copyright © 2025 The Authors. Published by Elsevier Masson SAS.. All rights reserved. DOI: 10.1016/j.biopha.2025.118558 PMID: 40972400 [Indexed for MEDLINE] Conflict of interest statement: Declaration of Competing Interest The authors declare no conflicts of interest related to the content of this manuscript. All authors have reviewed and approved the final version of the manuscript and confirm that they have no financial or personal relationships that could have inappropriately influenced or biased the work presented.
实验设计与方法 (Experimental Design & Methods)
采用结构生物学、计算机模拟和实验验证相结合的方法，系统分析蛋白质结构和功能关系。通过分子对接、动力学模拟等技术预测药物-靶点相互作用。
实验结果 (Experimental Results)
基于结构设计的小分子抑制剂活性提高10倍以上，成功解析了多个重要蛋白质的三维结构，为药物设计提供了结构基础。
数据汇总 (Data Summary)
基于结构设计的小分子抑制剂活性提高10倍以上，成功解析了多个重要蛋白质的三维结构，为药物设计提供了结构基础。
结论 (Conclusions)
基于蛋白质的药物研发策略为创新药物开发提供了新方向。
实践意义 (Practical Significance)
对推动靶向药物研发和精准医疗发展具有重要科学价值。