从单个浆细胞和B细胞中扩增IgG重链可变区（VH）和轻链可变区（VL，即Fab）
Amplification of IgG VH and VL (Fab) from single human plasma cells and B cells.
作者信息
Coronella J A, Telleman P, Truong T D, Ylera F, Junghans R P
机构信息
Biotherapeutics Development Lab, Harvard Institute of Human Genetics, Harvard Medical School and Beth Israel Deaconess Medical Center, Boston, MA 02215, USA.
出版信息
Nucleic Acids Res. 2000 Oct 15;28(20):E85. doi: 10.1093/nar/28.20.e85.
Abstract
Amplification of human immunoglobulin has many potential applications such as analysis of clonality, isolation of immunogenic antigens and antigen-specific immunotherapy. Here we describe a method for amplification of human immunoglobulin heavy and light chains from single B lymphocytes or plasma cells. Cells are isolated by FACS, and Ig is amplified by semi-nested RT-PCR. The method is versatile, sensitive and reliable: it provides appropriately paired heavy and light chains, requiring as little as 2 days to produce amplified Fab DNA from human tissues.
摘要
人免疫球蛋白的扩增有许多潜在应用，如克隆性分析、免疫原性抗原的分离和抗原特异性免疫治疗。在此，我们描述一种从单个B淋巴细胞或浆细胞中扩增产人免疫球蛋白重链和轻链的方法。通过荧光激活细胞分选术（FACS）分离细胞，并用半巢式逆转录聚合酶链反应（RT-PCR）扩增免疫球蛋白。该方法用途广泛、灵敏且可靠：它能提供适当配对的重链和轻链，从人组织中产生扩增的Fab DNA只需短短2天时间。
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