Single-domain antibody screening by isPLA-seq.
Single-domain antibody screening by isPLA-seq.
复制标题
通过 isPLA-Seq 筛选单域抗体
DOI：
10.26508/lsa.202101115
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发表时间：
2022-01
影响因子：
4.4
通讯作者：
Ma Z
中科院分区：
文献类型：
--
作者：
Yin Y;Yan F;Zhou R;Li M;Ma J;Liu Z;Ma Z
本研究描述了一种高灵敏度、高通量的单域抗体库筛选方法，该方法适用于通过isPLA-seq以单细胞分辨率筛选任何给定的感兴趣蛋白。单域抗体（sdAb）是一种具有广泛应用前景的研究策略。在这里，我们描述了一种方法，用于调整原位邻位连接测定（isPLA），然后进行测序（isPLA-seq），以便于以亚细胞和单细胞分辨率筛选给定蛋白质的高灵敏度，高通量sdAb文库。基于互补决定区3（CDR 3）的序列，重组sdAb可以被生产用于体外和体内应用。该方法为鉴定sdAb及其互补表位的功能及其在研究细胞过程中的潜在应用提供了一种通用手段。
This study describes a high-sensitive, high-throughput single-domain antibody library screening approach, which is applicable for any given interested protein at single-cell resolution by isPLA-seq. Single-domain antibody (sdAb) holds the promising strategies for diverse research and translational applications. Here, we describe a method for the adaptation of the in situ proximity ligation assay (isPLA) followed by sequencing (isPLA-seq) to facilitate screening of a high-sensitive, high-throughput sdAb library for a given protein at subcellular and single-cell resolution. Based on the sequence of complementarity-determining region 3 (CDR3), the recombinant sdAb can be produced for in vitro and in vivo utilities. This method provides a general means to identify the functional measure of sdAb and its complementary epitopes and its potential applications to investigate cellular processes.