[腐食酪螨过敏原的鉴定与纯化]

[Identification and purification of Tyrophagus putrescentiae allergens].

作者信息

Huang Zhi-Jian, Liu Zhi-Gang

机构信息

Allergy and Immunology Institute, Shenzhen University, Shenzhen 518060, China.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2007 Dec;25(6):483-7.

PMID:18441897

Abstract

OBJECTIVE

To identify and purify the allergens of Tyrophagus putrescentiae.

METHODS

Tyrophagus putrescentiae extract was analyzed by SDS-PAGE and Western blotting, and was partially purified by DE52 anion-exchange chromatography and HiLoad 16, 60 Superdex 200 prep grade Size-exclusion chromatography.

RESULTS

23 bands were found after SDS-PAGE with comparative molecular weight (Mr) of 177,000, 118,000, 107,000, 70,000, 67,000, 60,000, 52,000, 45,000, 41,000, 40,000, 38,000, 37,000, 35,000, 27,000, 23,000, 22,000, 18,000, 17,000, 16,000, 15,000, 14,000, 13,000 and 12,000. Five allergens were detected by Western blotting with Mr 128,000, 67,000-70,000, 36,000-37,000, 18,000 and 16,000, respectively. The positive reaction rate of 3 allergens, with Mr 128,000, 67,000-70,000 and 36,000-37,000, were 100%, while that of other 2 allergens with Mr 18,000 and 16,000 was 77.8% and 44.4% respectively. The allergen with Mr 18,000 was purified by anion-exchange chromatography and Size-exclusion chromatography.

CONCLUSION

Four major allergens and one minor allergen from Tyrophagus putrescentiae have been identified.

摘要

目的

鉴定并纯化腐食酪螨变应原。

方法

采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质免疫印迹法（Western blotting）分析腐食酪螨提取物，并通过DE52阴离子交换层析和HiLoad 16/60 Superdex 200预装级分子筛层析对其进行部分纯化。

结果

SDS-PAGE后发现23条带，相对分子质量（Mr）分别为177,000、118,000、107,000、70,000、67,000、60,000、52,000、45,000、41,000、40,000、38,000、37,000、35,000、27,000、23,000、22,000、18,000、17,000、16,000、15,000、14,000、13,000和12,000。蛋白质免疫印迹法检测到5种变应原，Mr分别为128,000、67,000 - 70,000、36,000 - 37,000、18,000和16,000。Mr为128,000、67,000 - 70,000和36,000 - 37,000的3种变应原阳性反应率为100%，而Mr为18,000和16,000的另外2种变应原阳性反应率分别为77.8%和44.4%。Mr为18,000的变应原经阴离子交换层析和分子筛层析纯化。