通过毛细管电泳分析大鼠气道表面液体微量样本中的蛋白质。
Analysis of proteins in microsamples of rat airway surface fluid by capillary electrophoresis.
作者信息
Govindaraju K, Cowley E A, Eidelman D H, Lloyd D K
机构信息
Montreal Chest Institute Research Centre, Royal Victoria and Montreal General Hospitals, McGill University, Quebec, Canada.
出版信息
J Chromatogr B Biomed Sci Appl. 1998 Feb 13;705(2):223-30. doi: 10.1016/s0378-4347(97)00508-2.
A thin layer of airway surface fluid (ASF) lining the pulmonary airways plays an important role in the primary defense mechanisms of the lung against bacterial infection. However, little is known about the composition of ASF due to the thinness (typically 5-30 microm in healthy animals) of the fluid layer and its relative inaccessibility, which causes considerable difficulties in sample collection and subsequent analysis. We have used a novel technique of capillary sampling coupled with capillary electrophoresis (CE) to analyze the protein composition of rat ASF. CE analyses were performed under two different conditions: a borate buffer, pH 9.1, or a phosphate buffer, pH 2.5, with 0.5 mM spermine. The different selectivities afforded by the two methods aid in peak identification, and quantitation of most of the major species was possible using both separation conditions. Albumin, transferrin and globulins are observed to be the major protein components in rat ASF, at concentrations of 28 mg ml(-1), 4.0 mg ml(-1) and 34 mg ml(-1) respectively, in comparison to 31 mg ml(-1), 3.1 mg ml(-1) and 40 mg ml(-1), respectively, in rat plasma.
覆盖在肺气道表面的一层薄薄的气道表面液体（ASF）在肺部抵御细菌感染的主要防御机制中发挥着重要作用。然而，由于该液体层很薄（在健康动物中通常为5 - 30微米）且相对难以获取，关于ASF的成分了解甚少，这给样本采集及后续分析带来了相当大的困难。我们采用了一种结合毛细管电泳（CE）的新型毛细管采样技术来分析大鼠ASF的蛋白质组成。CE分析在两种不同条件下进行：pH 9.1的硼酸盐缓冲液，或pH 2.5且含有0.5 mM精胺的磷酸盐缓冲液。两种方法提供的不同选择性有助于峰的识别，并且在两种分离条件下都能够对大多数主要成分进行定量分析。观察到白蛋白、转铁蛋白和球蛋白是大鼠ASF中的主要蛋白质成分，其浓度分别为28 mg ml⁻¹、4.0 mg ml⁻¹和34 mg ml⁻¹，而大鼠血浆中相应的浓度分别为31 mg ml⁻¹、3.1 mg ml⁻¹和40 mg ml⁻¹。
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