4种静注人免疫球蛋白制品的自身IgG抗体多样性研究

Diversity of IgG autoantibodies of 4 Chinese intravenous immunoglobulins

目的建立静注人免疫球蛋白(intravenous immunoglobulin,IVIG)制品自身IgG抗体谱展示方法及抗体种类分析方法,考察比较国内不同血液制品企业市售IVIG自身IgG抗体多样性。方法收集我国4个血液制品企业IVIG为研究对象,以人脐静脉内皮细胞总蛋白为抗原,通过双向电泳结合蛋白印记方法展示不同IVIG的自身抗体谱,通过IVIG与人类蛋白质组芯片杂交,高通量鉴定不同IVIG中自身抗体的种类并进行生物信息学分析。结果建立了IVIG自身抗体谱展示方法和抗体种类分析方法,获得了较高质量的IVIG自身抗体谱以及其能够识别的人自身蛋白的生物学信息,不同IVIG制品识别自身抗原的数量、种类有明显差异。4个制品针对人脐静脉内皮细胞蛋白抗体数量分别为241~386个;识别人蛋白质芯片上蛋白数量分别为292~435个,有172个人自身蛋白被4个制品均识别。结论利用抗体谱展示和蛋白质芯片技术能够用于分析IVIG中自身IgG抗体数量及种类多样性,检测的4个厂家IVIG制品中均具有广谱自身抗体,且具有各自的独特性。

Objective To develop methods to display the IgG autoantibody repertoire of intravenous immunoglobulin(IVIG)products,analyze the different types of antibodies and study the diversity of IgG autoantibody in 4 IVIG preparations from different Chinese manufacturers.Methods Two-dimensional gel electrophoresis and immunoblotting with human umbilical vein endothelial cell(HUVEC)proteins were used to demonstrate the IgG autoantibody repertoire and the human protein microarray with bioinformatics analysis was employed to profile the immune reactive autoantigens of the 4 IVIG preparations.Results The methods to showcase the autoantibody repertoire and study the antibody diversity of IVIG were successfully established.High-quality repertoires of IVIG autoantibodies and biological information about self-proteins that can be recognized were obtained.There was a significant difference in the recognition of the quantity and variety of the self-antigens by different IVIG products.The number of antibodies against HUVEC proteins in four products ranged from 241-386.The number of proteins recognized on the human protein chip ranged from 292-435,with 172 human self-proteins recognized by all four products.Conclusion Demonstration of antibody repertoire and protein chip technology can be used to analyze IVIG products′IgG autoantibody repertoire.All four preparations tested in this study exhibited a broad spectrum of antibodies against HUVEC proteins and human proteome microarray,each product had its unique antibody repertoire characteristics.