Issaq H J
Separation Technology Group, SAIC-Frederick/NCI-Frederick, MD 21702, USA.
Electrophoresis. 2001 Oct;22(17):3629-38. doi: 10.1002/1522-2683(200109)22:17<3629::AID-ELPS3629>3.0.CO;2-O.
In the last few years there has been an increased effort into the separation, quantification and identification of all proteins in a cell or tissue. This is a review of the role gel electrophoresis, high performance liquid chromatography (HPLC), and capillary electrophoresis (CE) play in proteomics research. The capabilities and limitations of each separation technique have been pointed out. Instrumental strategies for the resolution of cell proteins which are based on efficient separation employing either a single high-resolution procedure or a multidimensional approach on-line or off-line, and a mass spectrometer for protein identification have been reviewed. A comparison of the advantages of multi-dimensional separations such as two-dimensional polyacrylamide gel electrophoresis, HPLC-HPLC, and HPLC-CE to the separation of cell and tissue proteins are discussed. Also, a discussion of novel approaches to protein concentration, separation, detection, and quantification is given.
在过去几年中,人们加大了对细胞或组织中所有蛋白质进行分离、定量和鉴定的力度。本文综述了凝胶电泳、高效液相色谱(HPLC)和毛细管电泳(CE)在蛋白质组学研究中所起的作用。指出了每种分离技术的能力和局限性。综述了基于高效分离的细胞蛋白质解析的仪器策略,该策略采用单一高分辨率程序或在线或离线的多维方法,以及用于蛋白质鉴定的质谱仪。讨论了二维聚丙烯酰胺凝胶电泳、HPLC-HPLC和HPLC-CE等多维分离相对于细胞和组织蛋白质分离的优势。此外,还讨论了蛋白质浓缩、分离、检测和定量的新方法。