霜天蛾昆虫变应原多克隆抗体的制备及免疫特性分析

目的 制备兔抗霜天蛾变应原多克隆抗体,分析其与特异性过敏患者血清抗体成分的异同,为免疫筛选cDNA文库,制备基因重组霜天蛾变应原奠定基础.方法 应用霜天蛾变应原提取液免疫家兔获得霜天蛾变应原多克隆抗体,用酶联免疫吸附试验(ELISA)法测定混合免疫兔血清抗体效价:应用免疫印迹的方法对比分析免疫兔血清与特异性过敏患者血清抗体成分的异同.结果 混合免疫兔血清效价经ELISA检测,效价>1:10 000,免疫印迹检测兔抗血清免疫球蛋白G

Preparation and immune characteristic analysis of polyclonal antibody against Psilogramma menephron allergen

Objective To prepare polyclonal antibody of the rabbit anti-Psilogramma menephron allergen and analyse the similarities and differences of the antibody components about rabbit serum and patients serum with specific Psilogramma menephron allergy.The study provided the bases for immunoscreening Psilogramma menephron cDNA library and preparing the recombinant Psilogramma menephron allergen.Methods The polyclonal antibody was obtained by immunizing rabbits with Psilogramma menephron extracts.And its valence was tested with ELISA.The similarities and differences of the antibody components about rabbit serum and patients' serum with specific Psilogramma menephron allergy were tested with immunoblotting.Results The valence of mixed rabbits antiserum was more than 1:10 000 with ELISA.The lgG-immunoblotting analysis of rabbit antiserum showed that the protein bands of molecular mass were 100 000,92 000,79 000,74 000,66 000,49 000,43 000,36 000,25 000,16 000.The results of analyzing 9 patients'sera with allergic to Psilogramma menephron showed that the protein bands of molecular mass were 79 000,74 000,66 000,49 000,36 000,25 000.Conclusions The prepared rabbit antiserum recognized the similar protein bands of Psilogramma menephron allergen with specific allergic patients'serum and it could combine more IgG bands than patients'.The immunized rabbit serum can be used as probe for immunoscreening Psilogramma menephron cDNA library and be used in identifying the expressed recombinant allergen of Psilogramma menephron.