电喷雾-差分电迁移率-粒子计数法绝对定量分析牛血清白蛋白

Absolute Quantification of Bovine Serum Albumin by Electrospray-Differential Mobility Analysis-Condensation Particle Counter

建立了基于电喷雾-差分电迁移率-粒子计数的蛋白定量分析方法,通过优化电喷雾电压、液体溶液流速以及气体流速等参数,对牛血清白蛋白标准物质进行了定量分析。结果表明,在泰勒锥状态下,样品溶液流速300 nL/min,空气流速1.0 L/min,二氧化碳流速0.1 L/min的条件下,可以实现牛血清白蛋白标准物质和溶液标准物质的准确定量分析,测定结果为分别为20.05 g/L和69.52 g/L,与标准物质定量值的相对误差在±5%以内。该方法快速准确、灵敏度高、耗样量少,且不需要内标和外标,为蛋白质绝对定量分析提供了一种新的方法。

A method was established for the quantitative analysis of protein based on electrospray-differential electromigration-particle counting.Effects of factors such as electrospray voltage,liquid solution flow rate and gas flow rate on particle size distribution were systematically investigated and optimized,and common silica capillaries with an inner diameter of 25 μm and a length of 4.07 cm were used.The method was applied to the quantitative analysis of bovine serum albumin reference materials.Results showed that,under the condition of Taylor cone,a bovine serum albumin reference material and a bovine serum albumin solution reference material could both be accurately quantified at a sample solution flow rate of 300 nL/min,an air flow rate of 1.0 L/min and a carbon dioxide flow rate of 0.1 L/min,with the determined results of 20.05 g/L and 69.52 g/L,respectively.The relative errors were within ±5%,which were consistent with the quantitative values of standard materials.With no need of internal and external standards,the method is fast,accurate and sensitive,and it provides a new analytical approach for absolute protein quantification.