重组扁豆过敏原 Len c 1 表达纯化及免疫活性鉴定.

Objective: To purify recombinant lentils allergens Len c1 (rLen c 1) and identify the immune activity. Methods: By means of prokaryotic expression system rLen c 1 was produced , then the target protein with Strep Ⅱ tag was purified with the method of affinity chromatography. BALB/c mice were randomly divided into control group (injected by normal saline) and allergen sensitized group (injected by rLen c 1). The mice were immuned by intraperitoneal injection to set up BALB/c mice lentils allergic model, then the total IgE and allergen specific IgE were detected by indirect ELISA to identify the immune activity of recombinant lentil allergen. Results: Len c 1 protein expression was successfully induced by IPTG，which was expressed in the form of inclusion body; with dialysis renaturation and affinity chromatography,the highly purified renatured protein rLen c 1 was gained, and mice sensitization model was established successfully. Compared with the control group of mice, the TIgE and allergen specific IgE significantly increased in sensitized group of mice. Conclusions: Purified rLen c 1 allergen protein with immune activity was obtained to lay a foundation for the preparation of monoclonal antibodies, clinical diagnosis and immunotherapy. [ABSTRACT FROM AUTHOR]