摘要
为获得有生物活性的FcεRIα亚基细胞外区及多克隆抗体 ,并探知其功能 ,构建FcεRIα亚基的细胞外区的原核表达载体PBAD/gIIIA/FcεRIα,经表达、纯化后用斑点杂交法检测其活性。纯化制品免疫兔子获得抗血清 ,并研究抗血清对FcεRI的作用。结果发现 ,经原核表达出的FcεRIα亚基的细胞外区能与IgE结合 ;获得具有特异性抗FcεRI的抗血清 ,可能抑制嗜碱粒细胞脱颗粒。实验提示原核表达系统能产生有生物学活性的FcεRIα亚基的细胞外区 ,用其制备的抗血清能够识别FcεRIα亚基的细胞外区 ,可能抑制嗜碱粒细胞脱颗粒 ,为研究FcεRIα表达调节。
To search for the extracellular domain of human FcεRIα subunit in order to obtain a specific antiserum with biological activities,the prokaryotic expression plasmid PBAD/gIIIA/ FcεRIα for genes encoding the expression product was constructed.After expression and purification the biological activities of the expression products were determined by dot-blotting technique.Then rabbits were immunized with this product to raise antibodies and the effect of the specific antiserum on the FcεRI was to be analyzed. The experimental results showed that the expressed extracellular domain of FcεRIα subunit could bind with IgE specifically,and its antiserum could block the degranulation of basophils. These results indicate that the extracellular domain of FcεRIα subunit alone is sufficient for the IgE binding with higher avidity and specificity. The preparation the specific antiserum against this protein may be useful for the study the pathogenesis of allergic diseases.
出处
《现代免疫学》
CAS
CSCD
北大核心
2004年第2期161-163,168,共4页
Current Immunology
基金
默沙东哮喘基金 2 0 0 0年
