Lowance Center for Human Immunology, Division of Rheumatology, Department of Medicine, Emory University School of Medicine, Atlanta, GA, USA.
Department of Medicine, Division of Rheumatology, Emory University, Atlanta, GA, USA.
Methods Mol Biol. 2022;2421:231-241. doi: 10.1007/978-1-0716-1944-5_16.
Repertoire sequencing of B cells is the high-throughput profiling of B cell receptors (BCR) expressed on the surface of B cells and of immunoglobulins (Ig) expressed by antibody secreting cells. Each BCR/Ig transcript has a unique complementarity-determining region 3 (CDR3) sequence that can be used to identify and track individual B cell lymphocytes over time and throughout different compartments of the human body. B cell differentiation can be further tracked by assessing the point mutations acquired during affinity maturation via somatic hypermutation (SHM). Here we describe a method for high-throughput sequencing of the variable region of Ig heavy-chain transcripts for repertoire analysis of human B cells on the Illumina Miseq platform.
B 细胞受体（BCR）和分泌抗体的细胞表达的免疫球蛋白（Ig）的高通量分析是 B 细胞的序列分析。每个 BCR/Ig 转录本都有一个独特的互补决定区 3（CDR3）序列，可用于识别和跟踪随时间推移和在人体不同部位的单个 B 细胞淋巴细胞。通过评估体细胞高频突变（SHM）过程中获得的点突变，还可以进一步跟踪 B 细胞分化。在此，我们描述了一种在 Illumina Miseq 平台上用于人类 B 细胞免疫球蛋白重链可变区的高通量测序的方法，用于进行 B 细胞受体库分析。
