BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3601

Peptides:

Sequences	Affinity Values
TWWNPRLVYFDY(32) GVWWTWGNYGQM(5) YTPNWHFRWMPA(2) APTTWFNSDSIT(2)	[248.506±48.1295] [99.1517±25.2107] [82.7265±48.3205] [13.5881±14.3243]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (common panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 34962367

Target Name:

Graphene paper, GP

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-12 phage display library (X12)

Affinity Measurement Method:

Affinity Measurement Description:

To assess the binding affinity of GP-binding peptides, four selected GP-binding phages (with wild-type M13 phage as a control group) were amplified by infecting E. coli. After GP containers were UV treated and washed by TBS buffer, 200 μL of the corresponding phage solution (1010 pfu/ mL) was added into five containers separately and incubated for 60 min. The unbound phages were washed away with TBST. To get the GP-bound phages out of the GP substrates, 240 μL of the elution buffer was allowed to interact with the substrates for 8 min. Finally, 40 μL of neutralizer was added, and 10 μL of resultant was titrated to quantify the phages.

Brief Description:

First, GP was folded into a container and exposed to ultraviolet irradiation for an hour. Then, the container was washed using TBS. The phage library (2.0e11 pfu) was added to the GP container for incubation. After 1 h of incubation, the unbound phages were washed away with TBST. To elute bound phages, 240 μL of glycine-HCl (0.2 M, pH 2.2) was added and incubated for 8 min. Then the eluate was neutralized with 40 μL of Tris-HCl (1 M, pH 9.1). Then, 10 μL of the eluate was titrated. The rest of the eluate was amplified in ER2738 E. coli culture and treated as a sublibrary for the next round of biopanning. Output phages in round 3 were sequenced to identify the sequences of foreign peptides displayed at their protein 3 (p3) tips.

BiopanningDataSet Comments:

We first screened a high-affinity graphene paper (GP) binding peptide (TWWNPRLVYFDY) by the phage display technique. Then we chemically conjugated the GP-binding peptide to the synthetic hydroxyapatite (HA) nanorods. The GP-binding peptide on the resultant HA nanorods enabled them to be bound and assembled onto the GP substrate with high affinity, forming a GP-peptide-HA composite with significantly improved hydrophilicity of GP. The composite promoted the attachment and proliferation of mesenchymal stem cells (MSCs), demonstrating its outstanding biocompatibility. Due to the unique compositions of the composite, it was also found to induce osteogenic differentiation of MSCs in vitro in the absence of other inducers in the medium, by verifying the expression of the osteogenic markers including collagen-1, bone morphogenetic proteins 2, runx-related transcription factor 2, osteocalcin, and alkaline phosphatase. Our work suggests that the GP-binding peptide can be used to link inorganic nanoparticles onto GP to facilitate the biomedical applications of GP.

Curator:

Bowen Li

Curation Date:

2023-10-13

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