Jeanroy Frédéric, Gil Julie, Comby-Zerbino Clothilde, Demesmay Claire, Dugas Vincent
Universite Claude Bernard Lyon 1, ISA, UMR 5280, CNRS, 5 Rue de la Doua, 69100 Villeurbanne, France.
Universite Claude Bernard Lyon 1, ILM, UMR 5306, CNRS, 69100 Villeurbanne, France.
Molecules. 2024 Nov 28;29(23):5642. doi: 10.3390/molecules29235642.
The study of biomolecules and their interactions in their natural environment requires increasingly sophisticated technological and methodological developments. The complexity of these developments is due, among other things, to the nature of these molecules and the small quantities available depending on their origin. In this context, this study focuses on the conditions for improving the detection of glycosaminoglycans on a miniaturized scale by mass spectrometry. These multicharged anionic linear polysaccharides are in fact difficult to study by mass spectrometry and can present, for a given molecule, a large number of signals linked to different charge states, to the loss of one or more sulfate groups and to the presence of different adducts, which reduces sensitivity and complicates the interpretation of the spectra. In order to reduce this complexity, we have investigated different sample preparation methods applicable to small sample volumes. The development of home-made capillary ion-exchange columns, for example, makes it possible to control the adducts formed in nano-ESI coupling. However, their use on a miniaturized scale for detection by MALDI-TOF-MS does not allow for performances as high as those obtained with treatment with a commercial DOWEX resin. However, experimental results allowed us to demonstrate that the presence of DOWEX resin colloid residues in the aqueous phase greatly improves the quality of the spectra obtained by MALDI-TOFMS on a Fondaparinux model glycosaminoglycan.
对生物分子及其在自然环境中的相互作用进行研究，需要日益复杂的技术和方法发展。这些发展的复杂性，除其他因素外，还归因于这些分子的性质以及取决于其来源的少量可用量。在此背景下，本研究聚焦于通过质谱在小型化规模上改善糖胺聚糖检测的条件。这些多电荷阴离子线性多糖实际上难以通过质谱进行研究，并且对于给定分子，可能呈现大量与不同电荷状态、一个或多个硫酸根基团的丢失以及不同加合物的存在相关的信号，这降低了灵敏度并使光谱解释复杂化。为了降低这种复杂性，我们研究了适用于小体积样品的不同样品制备方法。例如，自制毛细管离子交换柱的开发使得能够控制在纳升电喷雾电离（nano-ESI）耦合中形成的加合物。然而，将其在小型化规模上用于基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）检测时，其性能不如用商业陶氏离子交换树脂（DOWEX树脂）处理所获得的性能高。然而，实验结果使我们能够证明水相中DOWEX树脂胶体残留物的存在极大地改善了在磺达肝癸钠模型糖胺聚糖上通过MALDI-TOF-MS获得的光谱质量。
