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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (06): 77-83    DOI: Q819
    
A study of SELDI-TOF-MS Protocol for Analyzing Proteins from Cells
MA Ke,WANG Qin,ZHANG Hong-gang,ZHANG Qiu-ju,XIU Rui-juan
Institute of Microcirculation,Peking Union Medical College & Chinese Academy of Medical Sciences,Beijing 100005,China
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Abstract  

Objective:To study the optimum protocol for analyzing proteins from cells by SELDI-TOF-MS.Methods:Proteins were extracted from cell samples by ultrasonic disruption, U9 cell disruption buffer and cell disruption buffer were established respectively. The concentrations of proteins were detected by BCA method, then protein samples were applied directly to the ProteinChips arrays using WCX magnetic bead and the bioprocessor, respectively, and protein samples were captured by Chips of WCX2, SAX2, IMACCu and H50, respectively, their results were compared, and detailed protein expression differences were detected using the chip of WCX2.Results:The concentrations of proteins isolated from cells using 3 different ways are: 0.25±0.034μg/μl，0.6±0.06μg/μl，1.02±0.077μg/μl,respectively. The method of Biochip Processor is simple, which requires less sample size and time. There is a good relationship between the protein peaks and protein concentrations in SELDI protein spectra. There are differences in the type of proteins captured by chips of WCX2, SAX2, H50, IMAC-Cu. In the limit of 1000~300 000Da, 87 protein expression difference peaks were detected by WCX2 chip, amount of which, 17 show a tendency variation.Conclusion:Using selfprepared cell disruption buffer receive highest protein concentration and which is better for further test.Bio-Chip Processor does better in application of sample.SELDI can not only detect protein expression differences but also reflect the protein concentration relationship; Selecting an appropriate chip or combining use of different chips to detect the protein expression differences can obtain more satisfactory results.

Key words： SELDI      Cell Proteins     
Received: 06 May 2010      Published: 12 June 2010
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Articles by authors
MA Ge
WANG Qin
ZHANG Gong-Gang
ZHANG Qiu-Ju
XIU Rui-Juan
Cite this article:

MA Ge, WANG Qin, ZHANG Gong-Gang, ZHANG Qiu-Ju, XIU Rui-Juan. A study of SELDI-TOF-MS Protocol for Analyzing Proteins from Cells. China Biotechnology, 2010, 30(06): 77-83.

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https://manu60.magtech.com.cn/biotech/Q819     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I06/77

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