硝酸纤维膜和丙酮沉淀尿蛋白质保存方法的比较

硝酸纤维膜法是一种简单、快速、经济的尿液蛋白质保存方法,但其与传统尿液蛋白质丙酮沉淀方法的差异有待进一步研究。相同尿液分别经硝酸纤维膜法和丙酮沉淀法制备尿蛋白质,经液相串联质谱分析鉴定蛋白质,采用谱图数定量,研究两种不同方法的差别。结果显示硝酸纤维膜法和丙酮沉淀法鉴定蛋白质数目几乎相同,鉴定蛋白质在谱图数的分布上几乎相同,鉴定蛋白质在蛋白质变异系数的分布上也几乎相同。因此,硝酸纤维膜法处理尿蛋白质与丙酮沉淀法基本一致,可以应用于大规模临床尿液样本的保存。

Comparison of two urinary protein preparation methods: nitrocellulose membrane preservation and acetone precipitation

Nitrocellulose membrane based urinary protein preservation method is simple, fast and economic, but its advantage over the traditionally used acetone precipitation method is still unclear. In this work, we prepared urinary proteins by the two methods by LC-MS/MS. Then we used protein spectra counts to assess the reproducibility of the two methods. Proteins identified by the two methods were almost the same in number, spectral count distribution and distribution of coefficients of variation value. In conclusion, nitrocellulose membrane method is generally the same as acetone precipitation method. It can be used for large scale preservation of clinical urine samples.