Merck
  • Home
  • Search Results
  • In vivo phage display: identification of organ-specific peptides using deep sequencing and differential profiling across tissues.

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

Nucleic acids research (2021-01-15)
Karlis Pleiko, Kristina Põšnograjeva, Maarja Haugas, Päärn Paiste, Allan Tobi, Kaarel Kurm, Una Riekstina, Tambet Teesalu
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Tannic acid, ACS reagent
Sigma-Aldrich
TWEEN® 20, for molecular biology, viscous liquid
Supelco
Tris(2-carboxyethyl)phosphine hydrochloride solution, 0.5 M, pH 7.0(aqueous solution; pH was adjusted with ammonium hydroxide)