Institute of Biomedicine, Research Centre for Integrative Physiology, and Pharmacology, Faculty of Medicine, University of Turku, Turku, Finland.
Department of Biosciences, Cell Biology, Faculty of Science and Engineering, Åbo Akademi University, Turku, Finland.
Proteomics. 2024 Jul;24(14):e2300340. doi: 10.1002/pmic.202300340. Epub 2024 Jun 14.
The breast milk composition includes a multitude of bioactive factors such as viable cells, lipids and proteins. Measuring the levels of specific proteins in breast milk plasma can be challenging because of the large dynamic range of protein concentrations and the presence of interfering substances. Therefore, most proteomic studies of breast milk have been able to identify under 1000 proteins. Optimised procedures and the latest separation technologies used in milk proteome research could lead to more precise knowledge of breast milk proteome. This study (n = 53) utilizes three different protein quantification methods, including direct DIA, library-based DIA method and a hybrid method combining direct DIA and library-based DIA. On average we identified 2400 proteins by hybrid method. By applying these methods, we quantified body mass index (BMI) associated variation in breast milk proteomes. There were 210 significantly different proteins when comparing the breast milk proteome of obese and overweight mothers. In addition, we analysed a small cohort (n = 5, randomly selected from 53 samples) by high field asymmetric waveform ion mobility spectrometry (FAIMS). FAIMS coupled with the Orbitrap Fusion Lumos mass spectrometer, which led to 41.7% higher number of protein identifications compared to Q Exactive HF mass spectrometer.
母乳成分包括多种生物活性因子，如活细胞、脂类和蛋白质。由于蛋白质浓度的动态范围较大且存在干扰物质，因此测量母乳血浆中特定蛋白质的水平具有挑战性。因此，大多数母乳蛋白质组学研究只能鉴定出不到 1000 种蛋白质。在乳蛋白质组研究中使用优化的程序和最新的分离技术，可以更精确地了解母乳蛋白质组。本研究（n=53）利用三种不同的蛋白质定量方法，包括直接 DIA、基于文库的 DIA 方法和直接 DIA 和基于文库的 DIA 相结合的混合方法。平均而言，我们通过混合方法鉴定了 2400 种蛋白质。通过应用这些方法，我们定量了与肥胖和超重母亲母乳蛋白质组相关的体重指数（BMI）变化。比较肥胖和超重母亲的母乳蛋白质组时，有 210 种蛋白质存在显著差异。此外，我们还通过高场非对称波形离子迁移谱（FAIMS）分析了一个小队列（n=5，从 53 个样本中随机选择）。FAIMS 与 Orbitrap Fusion Lumos 质谱仪联用，与 Q Exactive HF 质谱仪相比，蛋白质鉴定数量增加了 41.7%。
