两种链格孢霉变应原提取方法的比较  

Comparison of Novel and Traditional Methods for Alternaria Alternata Allergen Extract Production

作　　者：曹乃清[1] 张宏誊 杨秀云[1] 高鲁芳[1] 

机构地区：[1]山东大学 附属省立医院变态反应门诊,济南250021 [2]中国医学科学院 北京协和医学院 北京协和医院变态反应科,北京100730

出　　处：《中华临床免疫和变态反应杂志》2009年第1期21-24,共4页Chinese Journal of Allergy & Clinical Immunology

摘　　要：目的比较传统方法和新方法对链格孢霉变应原浸出液蛋白质含量、组分及生物活性的影响。方法将经空气曝皿获得、中国科学院鉴定为链格孢霉的菌株于26℃恒温下培养4周,取12份5 g湿重的菌苔,6份用传统方法(风干,高速粉碎机粉碎后,搅拌提取),6份采用新方法(液氮研磨+超声破碎,再搅拌提取)提取蛋白质;然后用改良的Bradford法进行蛋白质含量测定:SDS-PAGE分析两种方法提取的蛋白质组分的差别,放射性变应原吸附(RAST)抑制试验比较两者间变应原生物活性的差异。结果传统方法和新方法提取的蛋白质含量分别为(0.205±0.019)和(0.532±0.023)g/ml,所得到变应原的50%抑制率分别为5.96和1.25μg/ml。结论两种提取方法中,新方法能够得到较高的蛋白质含量及更多的蛋白质组分,对于低丰度蛋白的获得及生物学活性即效价的提高具有十分重要的意义。Objective To investigate the influence of novel and traditional methods on the extract production of Alternaria alternata. Methods Alternaria alternata was collected from the air and identified by Chinese Academy of Sciences, cultured under 26℃ for four weeks. Twelve 5g fresh molds, six extracted by traditional method （wind dry, disintegrated by high-speed disintegrator）, another six by novel method （liquid nitrogen＋ultrasonic）; protein concentration was determined by Bradford assay, its residual proteins were analyzed by SDS-PAGE, and bioactivitive potency were tested by RAST inhibition. Results The concentration of protein extractions were （0.205 ± 0.019） and （0.532 ± 0.023） g/ml respectively. From SDS-PAGE, we gained the most component in the extraction by novel method; from RAST, 50% inhibition were 5.96 and 1.25 μg/ml respectively. Conclusion Novel method was more production than traditional method.

关 键 词：传统方法 新方法 链格孢霉变应原浸出液 生物活性 放射性变应原吸附抑制试验 

分 类 号：R392.11[医药卫生—免疫学]