一种快速定量评估 B 细胞受体库 V 基因使用、特异性和克隆大小的方法。
A rapid and quantitative method for the evaluation of V gene usage, specificities and the clonal size of B cell repertoires.
机构信息
Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, United States.
出版信息
J Immunol Methods. 2012 Feb 28;376(1-2):143-9. doi: 10.1016/j.jim.2011.12.005. Epub 2011 Dec 28.
Abstract
The quantitative simultaneous description of both variable region gene usage and antigen specificity of immunoglobulin repertoires is a major goal in immunology. Current quantitative assays are labor intensive and depend on extensive gene expression cloning prior to screening for antigen specificity. Here we describe an alternative method based on high efficiency single B cell cultures coupled with RT-PCR that can be used for rapid characterization of immunoglobulin gene segment usage, clonal size and antigen specificity. This simplified approach should facilitate the study of antibody repertoires expressed by defined B cell subpopulations, the analysis of immune responses to self and nonself-antigens, the development and screening of synthetic antibodies and the accelerated study and screening of neutralizing antibodies to pathogenic threats.
摘要
定量同时描述免疫球蛋白库的可变区基因使用和抗原特异性是免疫学的主要目标。目前的定量检测方法劳动强度大，并且依赖于在筛选抗原特异性之前进行广泛的基因表达克隆。在这里，我们描述了一种基于高效的单细胞培养物与 RT-PCR 相结合的替代方法，可用于快速表征免疫球蛋白基因片段的使用、克隆大小和抗原特异性。这种简化的方法应该有助于研究特定 B 细胞亚群表达的抗体库、分析对自身和非自身抗原的免疫反应、合成抗体的开发和筛选以及对致病威胁的中和抗体的加速研究和筛选。
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