Comparison of Three Methods of Protein Extraction from Dermatophagoides Pteronyssinus for Two-dimensional Electrophoresis  

作　　者：Jin-lu Sun Hong-yu Zhang Zhi-yi Guo Wan-tao Ying Xiao-hong Qian Jing-lan Wang 

机构地区：[1]Department of Allergy, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences ＆ Peking Union Medical College, Beijing 100730, China [2]Experimental and Research Center, Key Laboratory of Occupational Health and Safety for Coal Industry, North China Coal Medical College, Tangshan, Hebei province 063000, China [3]Beijing Institute of Radiation Medicine, Beijing 100850, China

出　　处：《Chinese Medical Sciences Journal》2009年第1期64-68,共5页中国医学科学杂志（英文版）

基　　金：Supported by grants from the Ministry of Science and Technology of China (2008BAI59B04, 2003AA2Z3502);National Natural Science Foundation of China (30671943)

摘　　要：Obiective To explore an effective method of Dermatophagoides pteronyssinus protein extraction suitable for two-dimensional electrophoresis （2-DE） analysis. Methods The extracts of Dermatophagoides pteronyssinus were prepared with Coca＇s solution, lysis buffer of 2-DE, and Trizol reagent, respectively. Bicinchoninic acid （BCA） assay was used to determine the total protein concentration of the samples. The efficiency of different protein extraction methods were evaluated with 2-DE analysis. Results The concentrations of extracted protein by methods of Coca＇s solution, lysis buffer, and Trizol reagent were 0.63 g/L, 0.90 g/L, and 0.80 g/L, respectively. The 2-DE analysis results showed that some protein spots in low molecular weight （LMW） range could be detected with the Coca＇s solution method. With the lysis buffer of 2-DE method, more protein spots in LMW range could be detected, while the medium molecular weight （MMW） protein spots were absent. Several MMW protein spots （174-178 kD and 133 kD） and more LMW protein spots were detected withTrizol reagent method. Conclusions Among Coca＇s solution, lysis buffer of 2-DE, and Trizol reagent, the concentration of extracted protein of Dermatophagoides pteronyssinus by lysis buffer of 2-DE is the highest. However, most protein components of Dermatophagoides pteronyssinus purified mite bodies can be extracted by Trizol reagent, which may generally reflect the whole profile of Dermatophagoides pteronyssinus allergens.

关 键 词：house dust mite Dermatophagoides pteronyssinus ALLERGEN EXTRACTION two-dimensional electrophoresis 

分 类 号：R979.8[医药卫生—药品] R383.24[医药卫生—药学]