BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3528

Peptides:

Sequences
*CGGQDLKSC CSNLTSPC CPSSSREKC**

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (common panning)

Sequencing Platform:

MiSeq

Sequencing Method:

Next-generation sequencing

Rounds of Panning:

Reference:

PMID: 32243979

Target Name:

Cystic fibrosis mucus model

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

CX7C T7 phage display library (CX7C)

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

For the first round of selection, an initial phage amount of 4.2e9 phage plaque forming units (pfu) was added on top of the mucus layer in the donor compartment and incubated for 1 h at room temperature (25 °C). At 5, 10, 15, 30, and 45 min, 10 μL aliquots of the eluates were taken for further quantification. After 1 h, the entire eluate was collected from the basolateral side and titered using standard double layer plaque assay to quantify phage concentration. The eluted phage library was amplified in BL21 E. coli as described before, which was then quantified by plaque assay prior to the next round of selection. Two subsequent rounds of iterative selection were performed with initial amounts of 5.2e6 pfu and 4.2e9 pfu, respectively.

BiopanningDataSet Comments:

We used combinatorial peptide-presenting phage libraries and next-generation sequencing (NGS) to identify hydrophilic, net-neutral charged peptide coatings that enable penetration through human cystic fibrosis (CF) mucus ex vivo with ~600-fold better penetration than control, improve uptake into lung epithelial cells compared to uncoated or PEGylated-nanoparticles, and exhibit enhanced uniform distribution and retention in the mouse lung airways. These peptide coatings address multiple delivery barriers and effectively serve as excellent alternatives to standard PEG surface chemistries to achieve mucus penetration and address some of the challenges encountered using these chemistries. This biomolecule-based strategy can address multiple delivery barriers and hold promise to advance efficacy of therapeutics for diseases like CF.

Curator:

Chunchao Pu

Curation Date:

2020-12-29

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