Comprehensive monoclonal antibody generation

    公开(公告)号:US10670608B2

    公开(公告)日:2020-06-02

    申请号:US13977193

    申请日:2011-12-28

    申请人: Jay M. Short

    发明人: Jay M. Short

    摘要: The present invention relates to methods for efficiently generating recombinant monoclonal antibodies derived from B cells of a non-human host which has been immunochallenged with one or more target antigens. The methods comprise the steps of identifying and isolating B cell that bind to the antigen by FACS, and recombining and enriching for thousands of cells to create a B cell library. Related products and methods, such as methods of producing expression libraries, are also disclosed.

    GENE SITE SATURATION MUTAGENESIS
    4.
    发明申请
    GENE SITE SATURATION MUTAGENESIS 审中-公开
    基因座饱和度

    公开(公告)号:US20110165627A1

    公开(公告)日:2011-07-07

    申请号:US12817616

    申请日:2010-06-17

    申请人: Jay M. SHORT

    发明人: Jay M. SHORT

    IPC分类号: C12P19/34

    摘要: A method for producing progeny polynucleotides and polypeptides by Gene Site Saturation Mutagenesis (GSSM). The method provides a set of degenerate primers corresponding to codons of a template polynucleotide, and performs polymerase elongation to produce progeny polynucleotides, which contain sequences corresponding to the degenerate primers. The progeny polynucleotides can be expressed and screened for directed evolution.

    摘要翻译: 通过基因位点饱和诱变(GSSM)产生后代多核苷酸和多肽的方法。 该方法提供了一组对应于模板多核苷酸的密码子的简并引物,并进行聚合酶延伸以产生含有对应于简并引物的序列的后代多核苷酸。 可以表达和筛选后代多核苷酸用于定向进化。

    Capillary array-based enzyme screening
    10.
    发明授权
    Capillary array-based enzyme screening 有权
    基于毛细管阵列的酶筛选

    公开(公告)号:US06972183B1

    公开(公告)日:2005-12-06

    申请号:US09444112

    申请日:1999-11-22

    IPC分类号: C12N15/10 G01N33/573

    CPC分类号: C12N15/1086

    摘要: A process for screening an expression library to identify clones expressing enzymes having a desired activity is provided. The process involves first generating from genomic DNA samples of one or more microorganisms an expression library comprising a plurality of recombinant cell clones, and then introducing into capillaries in a capillary array a substrate and at least a subset of the clones, either individually or as a mixture. Interaction of the substrate and a clone expressing an enzyme having the desired activity produces an optically detectable signal, which can then be spatially detected to identify capillaries containing clones producing such a signal. The signal-producing clones can then be recovered from the identified capillaries.

    摘要翻译: 提供了筛选表达文库以鉴定表达具有所需活性的酶的克隆的方法。 该方法包括首先从一种或多种微生物的基因组DNA样品产生包含多个重组细胞克隆的表达文库,然后将毛细管阵列中的毛细管阵列引入基质和克隆的至少一个子集,单独地或作为 混合物。 底物和表达具有所需活性的酶的克隆的相互作用产生光学上可检测的信号,然后可以在空间上检测,以鉴定含有产生这种信号的克隆的毛细血管。 然后可以从识别的毛细血管中回收信号产生克隆。