Abstract:

In order to establish a technique system for differential proteomics of body fluids, cerebrospinal fluid samples in two different physiological states were studied. There was a Multile Sclerosis sample and a control for each group, which was done in triplicate. Samples were precipitated with ice-cold acetone and the protein concentration was accurately quantified. The control and abnormal groups were labeled with two different Cyanine Dyes (Cy2 and Cy5) using the minimal labeling method while the internal standard group was labeled with Cy3. Then 2D-E was used and three gels were run. Each gel contained a control group, an abnormal group and the internal standard group labeled with 3 different Cye Dyes. The information including differential express protein spots and their levels were obtained using a Typhoon 9400 scanner and DeCyder 2-D software. Protein spots were identified by MALDI-TOF／TOF. Metacore integrated software was used to analyze the interactions between proteins. The results show that a differential proteomics technique system is effective to discover valuable candidate proteins.

Key words: body fluids; differential proteomics; 2D-DIGE; Metacore