人IgE受体蛋白α片段免疫吸附剂的制备及效果评价  

Preparation and evaluation of an IgE-specific immunoadsorbent with recombinant FcεRIα

作　　者：张海珍 虢雅洁 傅俊成 陈校园 杨正根 ZHANG Hai-zhen;GUO Ya-jie;FU Jun-cheng;CHEN Xiao-yuan;YANG Zheng-gen(Guangdong Provincial Enterprise Key Laboratory for Blood Purification and Adsorption Technology,Guangzhou,Guangdong 510663,China;Guangzhou Koncen Bioscience Co.,Ltd.,Guangzhou,Guangdong 510660,China)

机构地区：[1]广东省血液净化吸附技术企业重点实验室,广东广州510663 [2]广州康盛生物科技股份有限公司,广东广州510660

出　　处：《热带医学杂志》2022年第11期1482-1486,共5页Journal of Tropical Medicine

基　　金：广东省科技计划项目(2020B121202021)

摘　　要：目的探讨人免疫球蛋白(Ig)E受体蛋白α片段免疫吸附剂对人血浆中IgE的吸附效果。方法构建含高亲和力受体α片段(FcεRIα)胞外区序列的重组真核表达载体pcDNA3.4,瞬时转染至HEK293F细胞,并用Ni亲和层析纯化蛋白。ELISA法测定受体蛋白与人IgE、IgG、IgM、IgA的亲和力,同时将受体蛋白偶联至琼脂糖上合成免疫吸附剂,测定吸附剂对人血浆中IgE的吸附性能。结果成功表达出重组受体蛋白,瞬时转染表达量为4 mg/L。纯化后蛋白分子量为30000 Mr,ELISA实验表明受体蛋白与人IgE的结合活性较高,是其与其它免疫球蛋白结合活性的7~8倍,交叉反应较小。将受体蛋白偶联至琼脂糖凝胶合成的吸附剂对IgE的吸附性能平均值为(10589±937)IU/mL,清除率平均值为(94.90±0.53)%。结论将受体蛋白α片段偶联至固相载体的免疫吸附剂可以有效地特异性吸附IgE,在治疗过敏性疾病方面前景广阔。Objective To explore the adsorption capacity of a immunoglobulin E(IgE)immunosorbent which was preparedby immobilizing the recombinant alpha segment of high affinity Ig E receptor(FcεRIα)on bestarose 6FF.Methods Recombinant plasmids pc DNA3.4 was constructed by genetic engineering.HEK293F was selected as expressing strain;recombinant protein was purified by Ni chromatography.The affinity and specificities with Ig E,Ig G,Ig M and Ig A weredetermined with ELISA.The recombinant FcεRIαwas immobilized on bestarose 6FF and the adsorption capacity for Ig E wasinvestigated by static adsorption.ResultsA fusion protein with molecular weight of 30000 Mr was expressed and purified.The expression of the FcεRIαwas about 4 mg/L.The binding activity of FcεRIαto human Ig E was 7-8 times higher thanthat of other immunoglobulins,such as Ig G,Ig M and Ig A;the cross-reaction was low.The FcεRIαcould bind Ig E withhigh affinity and with low cross-reactive with Ig G,Ig M and Ig A,which was evaluated with ELISA.The adsorption capacityof the adsorbent was about(10589±937)IU/m L,and the removal rate was(94.90±0.53)%.ConclusionTherecombinant human Ig E receptor,FcεRIa,could be applied as a specific ligand for preparing the adsorbent to specificallyremove Ig E,which would be a promising approach for the treatment of patients with high levels of Ig E and showingsymptoms of severe allergies.

关 键 词：高亲和力受体α片段 IGE 吸附 

分 类 号：R593.1[医药卫生—内科学]