PROTOCOL FOR IDENTIFYING AND ISOLATING ANTIGEN-SPECIFIC B CELLS AND PRODUCING ANTIBODIES TO DESIRED ANTIGENS
    8.
    发明申请
    PROTOCOL FOR IDENTIFYING AND ISOLATING ANTIGEN-SPECIFIC B CELLS AND PRODUCING ANTIBODIES TO DESIRED ANTIGENS 审中-公开
    用于鉴定和分离抗原特异性B细胞并将抗体产生于所需抗原的协议

    公开(公告)号:US20160033504A1

    公开(公告)日:2016-02-04

    申请号:US14776945

    申请日:2014-03-18

    IPC分类号: G01N33/569

    摘要: Methods of identifying antigen-specific antibody-secreting and antibody-forming cells, such as antigen-specific B cells, and methods for cloning the antigen-specific antibody sequences of the antibody produced by these cells are provided. In particular, the methods include enriching B cells for antigen-specific B cells, culturing the antigen-specific B cells to generate clonal B cell populations, detecting clonal B cells that produce a single antigen-specific antibody, optionally screening the clonal B cell populations for functional activity, staining and sorting the cells to isolate the antigen-specific B cells, sequencing the nucleic acids encoding the antigen-specific antibody sequences, expressing the sequences to produce an antibody, isolating the antibody and screening the antibody for antigen recognition. The methods provide improved enrichment and selection of antigen-specific antibody-secreting and antibody-forming cells, which enhances recovery of antigen-specific antibodies.

    摘要翻译: 提供了鉴定抗原特异性抗体分泌型和抗体形成细胞如抗原特异性B细胞的方法,以及用于克隆由这些细胞产生的抗体的抗原特异性抗体序列的方法。 特别地,所述方法包括富集B细胞用于抗原特异性B细胞,培养抗原特异性B细胞以产生克隆B细胞群体,检测产生单一抗原特异性抗体的克隆B细胞,任选筛选克隆B细胞群 用于功能活性,染色和分选细胞以分离抗原特异性B细胞,测序编码抗原特异性抗体序列的核酸,表达序列以产生抗体,分离抗体并筛选抗体以进行抗原识别。 该方法提供改进的抗原特异性抗体分泌和抗体形成细胞的富集和选择,其增强抗原特异性抗体的回收。