BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3554

Peptides:

Sequences
GFPSVRDLSPLR WATLDLGPQPYS PETWTHW HQSQSRM PLSKGKL NTSPLNT PTKWSAT ASGFTAT PNSVKAQ TMSTTQV GLELREK

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (in vivo)

Sequencing Platform:

Illumina MiSeq

Sequencing Method:

Next-generation sequencing

Rounds of Panning:

Reference:

PMID: 31589023

Target Name:

C57bl/6 mice spinal cord

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-12 and Ph.D.-C7C phage display library pool

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

For library screening, Ph.D.−12 and C7C phage library (New England Biolabs, USA) were mixed, 1.5e13 pfu/mL, and injected intraperitoneally (IP) into C57bl/6 mice (100 μl, n=2 per group). Prior to tissue harvest, mice were euthanized and perfused with 20 mL saline to ensure complete tissue exsanguination. Spinal cord, brain, and cardiac tissue was harvested and flash-frozen in DMEM (Invitrogen, Inc.) with 1% BSA (Fisher Scientific). After homogenization, tissue homogenates were centrifuged and resuspended in 1% Triton-X 100 (Sigma-Aldrich, Inc.) and passed through a 25-gauge needle to ensure cell lysis. Phage titer was quantified and spinal- and brain-tissue homogenate was amplified for subsequent screens.

BiopanningDataSet Comments:

The series of TACL peptides were synthesized and tested for their ability to deliver a model enzyme (NeutrAvidin-horseradish peroxidase fusion) to the brain and spinal cord. Three TACL-peptides facilitated significant active enzyme delivery into the CNS, with limited accumulation in off-target organs. Peptide structure and serum stability is increased when internal cysteine residues are cyclized by perfluoroarylation with decafluorobiphenyl, which increased delivery to the CNS further. TACL-peptide was demonstrated to localize in parasympathetic ganglia neurons in addition to neuronal structures in the hindbrain and spinal cord. By targeting uptake into ANS neurons, we demonstrate the potential for TACL-peptides to bypass the blood-brain barrier and deliver therapeutics into the brain and spinal cord.

Curator:

Chunchao Pu

Curation Date:

2020-12-29

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