Nägele Edgar, Vollmer Martin, Hörth Patric, Vad Cornelia
Agilent Technologies, R&D and Marketing GmbH & Co. KG, Hewlett-Packard-Str. 8, Waldbronn, Germany.
Expert Rev Proteomics. 2004 Jun;1(1):37-46. doi: 10.1586/14789450.1.1.37.
Today, 2D online or offline liquid chromatography/mass spectrometry is state of the art for the identification of proteins from complex proteome samples in many laboratories. Both 2D liquid chromatography methods use two orthogonal liquid chromatography separation techniques. The most commonly used techniques are strong cation exchange chromatography for the first dimension and reversed phase separation for the second dimension. In order to improve sensitivity the reversed phase separation is usually performed in the nanoflow scale and mass spectrometry is used as the final detection method. The high-performance liquid chromatography techniques complement the 2D-gel techniques supporting their weaknesses. This is especially true for the gel separation of hydrophobic membrane proteins, which play an important role in living cells as well as being important targets for future pharmaceutical drugs.
如今，二维在线或离线液相色谱/质谱联用技术是许多实验室从复杂蛋白质组样品中鉴定蛋白质的先进技术。两种二维液相色谱方法均采用两种正交的液相色谱分离技术。最常用的技术是一维的强阳离子交换色谱和二维的反相分离。为了提高灵敏度，反相分离通常在纳流规模下进行，质谱用作最终检测方法。高效液相色谱技术弥补了二维凝胶技术的不足。对于疏水性膜蛋白的凝胶分离尤其如此，疏水性膜蛋白在活细胞中起着重要作用，也是未来药物的重要靶点。
