BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3557

Peptides:

Sequences	Affinity Values
LLADTTHHRPWT HATGTHGLSLSH TSGYLHLRGRWR	[0.472097±0.001114,0.32025] [0.074187±0.001120,0.118277±0.001120] [0.03526,0.0615]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 31786302

Target Name:

C-terminal domain of CCN family member 2

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:

ELISA

Affinity Measurement Description:

Positive phage clones were adsorbed to PKW-CTGF- and TrxA-CTGF-coated microplates, and the optical density at 450 nm (OD450) was directly determined using ELISA. OD450 values were reproduced from Figure 2A and shown. The first column of affinity values was the binding affinity of phage clones to PKW-CTGF, and the second column the binding affinity of phage clones to TrxA-CTGF.

Brief Description:

Human TrxA-CTGF/C and TrxA dissolved in sodium bicarbonate (NaHCO3, pH 8.6) were coated onto 96-well plates and incubated overnight at 4 °C. After blocking with 0.5% (w/v) bovine serum albumin (BSA), the wells were washed with TBST 6 times. The random peptide library was added to the TrxA-coated plate and incubated at 37 °C for 1 h. Supernatants were then added to the TrxA-CTGF/C-coated plate for 1 h at 37 °C. The plates were then washed six times with TBST and eluted with 0.2 M glycine/HCl (pH 2.2) containing 0.5% BSA, then immediately neutralized with Tris/HCl (pH 9.1). Next, phages were inoculated into LB medium with 10% E2738 at 37 °C for 4.5 h and centrifuged at 30,000 ×g for 15 min at 4 °C, the supernatants added to 1/6 volume PEG8000/NaCl at 4 °C overnight. Four rounds of selection were performed as follows: the supernatants were centrifuged at 12,000 ×g for 15 min and suspended in 0.5% Tween20-TBST; in the second round washed 10 times 0.5% Tween20-TBST; in the third round, washed 15 times with 1% Tween20-TBST; and, finally, in the fourth round, washed 20 times with 2% Tween20-TBST.

BiopanningDataSet Comments:

810A (LLADTTHHRPWT) was developed as a novel peptide, which binds to the C-terminal domain of connective tissue growth factor (CTGF). It could effectively inhibit the proliferation, migration, and expression of TGF-β and α-SMA in cells pretreated with CTGF. In in vivo experiments using an animal model of BLM-induced pulmonary fibrosis, the peptide demonstrated similar effects, which effectively reduced the onset and development of pulmonary fibrosis in mice.

Curator:

Chunchao Pu

Curation Date:

2020-12-29

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