Optimization protocol for amyloid-β peptides detection in human cerebrospinal fluid using SELDI TOF MS

被引:28
作者
Albertini, Valentina [1 ]
Bruno, Anna [2 ]
Paterlini, Anna [1 ]
Lista, Simone [3 ]
Benussi, Luisa [2 ]
Cereda, Cristina [3 ]
Binetti, Giuliano [2 ]
Ghidoni, Roberta [1 ,2 ]
机构
[1] Ctr S Giovanni DioFBF, Prote Unit, IRCCS, Brescia, Italy
[2] Ctr S Giovanni DioFBF, Memory Clin, IRCCS, NeuroBioGen Lab, Brescia, Italy
[3] Univ Pavia, Neurol Inst C Mondino, IRCCS, Lab Expt Neurobiol, I-27100 Pavia, Italy
关键词
Alzheimer's disease; 6E10; and; 4G8; Cerebrospinal fluid; N and C-terminally truncated Amyloid-beta peptides; PS20 ProteinChip array; DOWNS-SYNDROME BRAINS; ALZHEIMERS-DISEASE; A-BETA; MASS-SPECTROMETRY; PATHOGENESIS; HYPOTHESIS; MUTATION; PLAQUES; PROTEIN;
D O I
10.1002/prca.200900166
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Purpose: The aim of the present work was to set up an optimized protocol for human cerebrospinal fluid amyloid-beta (A beta) profiling. Experimental design: We devised an immunoproteomic assay that employs monoclonal antibodies (mAbs) on Preactivated Surface (PS20) chip array followed by SELDI TOF MS. A comparison of a number of factors was performed, and the impact of these differences was noted. Each variable was tested using in parallel two different mAbs, 6E10 and 4G8. In addition, we tested whether the combined use of these two mAbs could improve the capture of N and C-terminally truncated A beta peptides and then the quality of spectra. Results: The best results were obtained using a mixture of A beta mAbs (0.125 mu g/mu L 6E10+4G8): 15 A beta peptides (including 3 N-terminally truncated forms) were detected. Conclusions and clinical relevance: This approach has many potential advantages in speed, sensitivity and economy of reagents and could be helpful in order to define the role played by specific A beta truncated forms in cognitive decline.
引用
收藏
页码:352 / 357
页数:6
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