Aims: This study aimed to generate an improved method of protein extraction and purification from plant tissues containing very high amounts of phenolic compounds and other interfering biomolecules.

Background: Protein extraction at proteomic studies on some plant species, including conifers, is challenging, and the yield and quality are unpredictable.

Objective: Two popular protocols were combined with each other to construct a novel one with enhanced abilities to produce higher purity of samples compatible for high precision molecular systems and analysis.

Methods: The new method was compared with the other two for their efficiencies in classical SDS- PAGE, 2-DE and capillary chromatography applications.

Results: All three methods were comparable in SDS-PAGE procedure; however, only the new method created acceptable gel images in 2-DE. Bioanalyzer results, also, demonstrated that the new method provided protein samples pure enough to be used in capillary chromatography with 2 times more peaks in electropherograms with lower noise and higher total relative protein concentrations closest to the applied amount.

Conclusion: The new combined method is a successful alternative for plant proteomicists with higher yield and quality of proteins from recalcitrant tissues.

Other: The new method could be preferred, especially, for high-tech, sensitive proteomic analysis.

目的：本研究旨在从含有大量酚类化合物和其他干扰生物分子的植物组织中产生一种改进的蛋白质提取和纯化方法。

背景：在一些植物物种（包括针叶树）的蛋白质组学研究中提取蛋白质具有挑战性，并且产量和质量是不可预测的。

目标：将两种流行的协议相互结合，构建一种新的协议，该协议具有增强的能力，可产生与高精度分子系统和分析兼容的更高纯度的样品。

方法：将新方法与其他两种方法在经典 SDS-PAGE、2-DE 和毛细管色谱应用中的效率进行了比较。

结果：所有三种方法在 SDS-PAGE 程序中具有可比性；然而，只有新方法在 2-DE 中创建了可接受的凝胶图像。生物分析仪的结果还表明，新方法提供的蛋白质样品纯度足以用于毛细管色谱，其电泳图谱中的峰多出 2 倍，噪音更低，总相对蛋白质浓度更高，最接近应用量。

结论：新的组合方法是植物蛋白质组学家的成功替代方法，可从顽固组织中获得更高的蛋白质产量和质量。

其他：新方法可能是首选，尤其是对于高科技、灵敏的蛋白质组学分析。