In vivo phage display: identification of organ-specific peptides using deep sequencing and differential profiling across tissues

Nucleic Acids Res. 2021 Apr 19;49(7):e38. doi: 10.1093/nar/gkaa1279.


In vivo phage display is widely used for identification of organ- or disease-specific homing peptides. However, the current in vivo phage biopanning approaches fail to assess biodistribution of specific peptide phages across tissues during the screen, thus necessitating laborious and time-consuming post-screening validation studies on individual peptide phages. Here, we adopted bioinformatics tools used for RNA sequencing for analysis of high-throughput sequencing (HTS) data to estimate the representation of individual peptides during biopanning in vivo. The data from in vivo phage screen were analyzed using differential binding-relative representation of each peptide in the target organ versus in a panel of control organs. Application of this approach in a model study using low-diversity peptide T7 phage library with spiked-in brain homing phage demonstrated brain-specific differential binding of brain homing phage and resulted in identification of novel lung- and brain-specific homing peptides. Our study provides a broadly applicable approach to streamline in vivo peptide phage biopanning and to increase its reproducibility and success rate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Surface Display Techniques / methods*
  • High-Throughput Nucleotide Sequencing / methods
  • High-Throughput Screening Assays / methods*
  • Mice
  • Mice, Inbred BALB C
  • Peptide Library*
  • Peptides / metabolism*
  • Tissue Distribution


  • Peptide Library
  • Peptides