单细胞 B 细胞测序生成天然配对的 scFab 酵母表面展示文库。
Single-Cell B-Cell Sequencing to Generate Natively Paired scFab Yeast Surface Display Libraries.
机构信息
Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI, USA.
Institute for Quantitative Health Sciences and Engineering, Michigan State University, East Lansing, MI, USA.
出版信息
Methods Mol Biol. 2023;2681:175-212. doi: 10.1007/978-1-0716-3279-6_11.
The immune cell profiling capabilities of single-cell RNA sequencing (scRNA-seq) are powerful tools that can be applied to the design of theranostic monoclonal antibodies (mAbs). Using scRNA-seq to determine natively paired B-cell receptor (BCR) sequences of immunized mice as a starting point for design, this method outlines a simplified workflow to express single-chain antibody fragments (scFabs) on the surface of yeast for high-throughput characterization and further refinement with directed evolution experiments. While not extensively detailed in this chapter, this method easily accommodates the implementation of a growing body of in silico tools that improve affinity and stability among a range of other developability criteria (e.g., solubility and immunogenicity).
单细胞 RNA 测序（scRNA-seq）的免疫细胞分析能力是一种强大的工具，可以应用于治疗性单克隆抗体（mAb）的设计。该方法使用 scRNA-seq 来确定免疫小鼠天然配对的 B 细胞受体（BCR）序列作为设计的起点，概述了一种简化的工作流程，用于在酵母表面表达单链抗体片段（scFabs），以进行高通量表征，并通过定向进化实验进一步改进。虽然本章没有详细介绍，但该方法很容易适应越来越多的计算工具的实施，这些工具可以提高亲和力和稳定性，同时满足其他一系列可开发性标准（例如溶解度和免疫原性）。
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