BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3542

Peptides:

Sequences

Affinity Values

VVSPDMNLLLTN(5/16)
VFSSMVHVLNTH(3/16)
SGVYKVAYDWQH(1/16)
GLHTSATNLYLH(1/16)
CYAGHDLYVAAD(1/16)
SLSWLTKMQMEM(1/16)

[0.912840±0.000310]
[0.140007±0.000179]
[0.235673±0.002483]
[1.061523±0.002488]
[NA]
[NA]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (common panning)

Sequencing Platform:

M13 DNA isolation Kit (Biomiga. Inc)

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 31993846

Target Name:

Escherichia coli O157:H7 cells

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:

The color development was recorded using a microplate reader (Thermo Fisher, MultiskanTM GO) by monitoring absorbance at 405 nm. A405 shown was reproduced from Figure 2.

Brief Description:

Briefly, 10 μL phage library (~e13 pfu/mL) were added into 1 mL E. coli O157:H7 cell suspensions (OD600 = 0.5) and incubated for 1 h at room temperature with gentle agitation. Bacteria with bound phages were precipitated by spinning for 5 min at 16,000×g, and separated from unbound phages in solution by a series of 10 washing and centrifugation steps (16,000×g, 5 min) with 1 mL TBST buffer (50 mM Tris–HCl, 0.05% (v/v) Tween 20) each time. After washing, bound phages with E. coli O157:H7 were suspended in 200 μL elution buffer (0.2 M glycine–HCl, pH 2.2) with gentle shaking at room temperature for 10 min. The eluted phages were neutralized with 150 μL, 1 M Tris–HCl (pH 9.1).

BiopanningDataSet Comments:

A 12-mer peptide (with the sequence of GLHTSATNLYLH) with high binding affinity toward Escherichia coli O157:H7 was discovered by biopanning of phage-displayed peptide library. The peptide modified with glycine residues (G3) and one cysteine (C) residue at C-terminal, could self-assemble on gold electrodes, enabling electrochemical impedance spectroscopy (EIS) analysis for quantitative detection of E. coli O157:H7. This method showed a low detection limit of 20 CFU/mL and a liner range from 2e2 to 2e6 CFU/mL.

Curator:

Chunchao Pu

Curation Date:

2020-12-29

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