Objective 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 with Trizol 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.