Single-domain antibody screening by is PLA-seq

Life Sci Alliance. 2021 Oct 21;5(1):e202101115. doi: 10.26508/lsa.202101115. Print 2022 Jan.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cell Line
  • Complementarity Determining Regions / chemistry
  • Complementarity Determining Regions / immunology
  • Gene Library
  • High-Throughput Screening Assays / methods*
  • Humans
  • Immunophenotyping
  • Molecular Imaging
  • Sequestosome-1 Protein / immunology
  • Single-Domain Antibodies / chemistry
  • Single-Domain Antibodies / immunology*

Substances

  • Complementarity Determining Regions
  • SQSTM1 protein, human
  • Sequestosome-1 Protein
  • Single-Domain Antibodies