CAJ | 학술논문

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<IgG)可以特异性识别霜天蛾10种蛋白组分,相对分子质量分别为100 000、92 000、79 000、74 000、66 000、49 000、43 000、36 000、25 000和16 000;患者血清IgG可以识别6种蛋白组分,相对分子质量分别为79 000、74 000、66 000、49 000、36 000和25 000.结论制备出的兔抗血清与特异性霜天蛾过敏患者血清特异性识别霜天蛾变应原的结果接近.比患者可以识别更多的蛋白条带,可以应用免疫兔血清作为筛选霜天蛾cDNA文库的探针及重组霜天蛾变应原表达蛋白的鉴定分析.
목적 제비토항상천아변응원다극륭항체,분석기여특이성과민환자혈청항체성분적이동,위면역사선cDNA문고,제비기인중조상천아변응원전정기출.방법 응용상천아변응원제취액면역가토획득상천아변응원다극륭항체,용매련면역흡부시험(ELISA)법측정혼합면역토혈청항체효개:응용면역인적적방법대비분석면역토혈청여특이성과민환자혈청항체성분적이동.결과 혼합면역토혈청효개경ELISA검측,효개>1:10 000,면역인적검측토항혈청면역구단백G<IgG)가이특이성식별상천아10충단백조분,상대분자질량분별위100 000、92 000、79 000、74 000、66 000、49 000、43 000、36 000、25 000화16 000;환자혈청IgG가이식별6충단백조분,상대분자질량분별위79 000、74 000、66 000、49 000、36 000화25 000.결론 제비출적토항혈청여특이성상천아과민환자혈청특이성식별상천아변응원적결과접근.비환자가이식별경다적단백조대,가이응용면역토혈청작위사선상천아cDNA문고적탐침급중조상천아변응원표체단백적감정분석.
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.