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Published on: September 30, 2019
Selection of peptide ligands for human placental transcytosis systems using in vitro phage display
Saleem Basha1, Shruthi Vaidhyanathan, Giovanni M Pauletti
1James L. Winkle College of Pharmacy, University of Cincinnati, Cincinnati, OH, USA.
Fetal pharmacotherapy generally relies on nonspecific biodistribution of therapeutic agents to the unborn child following drug administration into the maternal circulation system. Physiologically, transfer of polar, high-molecular weight solutes across the placenta is facilitated by a specialized, vesicular transport mechanism termed transcytosis. To develop biotechnology-based drugs such as proteins, DNA, and siRNA as clinically effective therapeutics, transcytosis systems have been evaluated as a promising strategy to augment drug transfer across endothelial and epithelial barriers. Screening of random peptide libraries using phage display is a powerful technology to identify peptide sequences with high affinity for surface proteins on desired target cells. Here, we describe assembly of a diverse, cyclic heptapeptide library on the icosahedral T7 bacteriophage platform. This phage-displayed library of random peptides was used for functional in vitro screens across BeWo cell monolayers to identify peptide ligands that facilitate placental transcytosis of viral particles across this cell culture model of the human trophoblast barrier.
Fetal pharmacotherapy generally relies on nonspecific biodistribution of therapeutic agents to the unborn child following drug administration into the maternal circulation system. Physiologically, transfer of polar, high-molecular weight solutes across the placenta is facilitated by a specialized, vesicular transport mechanism termed transcytosis. To develop biotechnology-based drugs such as proteins, DNA, and siRNA as clinically effective therapeutics, transcytosis systems have been evaluated as a promising strategy to augment drug transfer across endothelial and epithelial barriers. Screening of random peptide libraries using phage display is a powerful technology to identify peptide sequences with high affinity for surface proteins on desired target cells. Here, we describe assembly of a diverse, cyclic heptapeptide library on the icosahedral T7 bacteriophage platform. This phage-displayed library of random peptides was used for functional in vitro screens across BeWo cell monolayers to identify peptide ligands that facilitate placental transcytosis of viral particles across this cell culture model of the human trophoblast barrier.
