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Cell-based methods for coupling protein interactions and binding molecule selection

机译:基于细胞的蛋白质相互作用与结合分子选择偶联方法

摘要

The invention relates to cell-based methods for diversifying, expressing and selecting binding molecules, e.g., antibodies, and target molecules to which they bind, all of which are expressed in the same cell. The target molecule can be a member of a ligand binding pair comprising a cell-surface expressed ligand binding receptor molecule and its cognate ligand, which interact within the cell. The methods provide retaining either the antibody or its target in a cell organelle as the site of binding and interaction. By performance of the methods, the binding or non-binding of the antibody to its target molecule within the cell produces a cell phenotype that is detectable at the cell surface via high throughput assays, e.g., flow cytometry. The methods are particularly useful for generating, recovering and providing antibodies that have optimal target molecule binding properties or activities for potential therapeutic use. Methods for generating diversity in such antibodies are also provided.
机译:本发明涉及用于多样化,表达和选择结合分子(例如抗体)和它们结合的靶分子的基于细胞的方法,所有这些都在同一细胞中表达。靶分子可以是包含在细胞内相互作用的细胞表面表达的配体结合受体分子及其同源配体的配体结合对的成员。该方法提供将抗体或其靶标保留在细胞器中作为结合和相互作用的位点。通过该方法的执行,抗体与其在细胞内的靶分子的结合或不结合产生可通过高通量测定法例如流式细胞术在细胞表面检测到的细胞表型。该方法对于产生,回收和提供具有最佳靶分子结合特性或活性以用于潜在治疗用途的抗体特别有用。还提供了在此类抗体中产生多样性的方法。

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