Department of Pathology and Laboratory Medicine, Marsico Lung Institute, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Methods Mol Biol. 2024;2763:51-59. doi: 10.1007/978-1-0716-3670-1_4.
Membrane-bound mucins constitute a large portion of the periciliary layer of lung epithelial surfaces, and thus play an important role in many aspects of innate defense. The biophysical and biochemical properties of the membrane-bound mucins have important implications for mucociliary clearance, viral penetration, and potential therapeutics delivered to the airway surface. Hence, isolating them and determining these properties is important in understanding airways disease and ultimately in developing treatments. Here, we describe a method using isopycnic centrifugation to enrich and isolate shed membrane-bound mucins from the washings of human bronchial epithelial cell cultures.
膜结合粘蛋白构成肺上皮表面纤毛层的很大一部分,因此在先天防御的许多方面发挥重要作用。膜结合粘蛋白的物理化学特性对粘液纤毛清除、病毒穿透和递送到气道表面的潜在治疗剂具有重要意义。因此,分离它们并确定这些特性对于理解气道疾病并最终开发治疗方法非常重要。在这里,我们描述了一种使用等密度离心从人支气管上皮细胞培养物的洗涤液中富集和分离脱落的膜结合粘蛋白的方法。