BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 2866

Peptides:

Sequences

Affinity Values

LPADEDHHFYWT(8)
LLQLTTTKRPIT(7)
QTHHTHLPIQRA(5)
TINSNTLNSTPP(5)
QKESSTHFMAIH(3)
SFPTMTENFYPR(3)
MHDASPQLYRGR(2)
MEDNDTHWARMA(2)
TMYEPTTTRSPA(1)
SVTAGMPNRSNR(1)
HLSVAKKPLHRP(1)
WPGLLGTTSPNI(1)
VHWPASQPVQIP(1)
HDHNTNDMTLNA(1)

[2.85 ± 0.33][12.82 ± 1.73]
[5.80 ± 0.52][22.39 ± 2.30]
[3.54 ± 0.25][11.46 ± 0.42]
[5.36 ± 0.99][10.10 ± 0.31]
[3.61 ± 0.86][NT]
[3.18 ± 0.63][NT]
[4.28 ± 0.85][NT]
[3.07 ± 0.13][NT]
[2.72 ± 0.28][NT]
[3.85 ± 0.76][NT]
[2.20 ± 0.24][NT]
[2.32 ± 2.46][NT]
[2.24 ± 0.26][NT]
[2.70 ± 0.08][NT]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 25886968

Target Name:

Esophageal squamous carcinoma cell Eca-109

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,Fluorescence

Affinity Measurement Description:

Evaluation of the binding selectivity of 60 phage clones were performed by cellular ELISA. Eca-109 and HEK293 cells were used in ELISA. The 96-well plates were measured at 450 nm using an ELISA reader (Bio-Tek ELX800, USA). Irrelevant phage clone (IRP, an amplified phage randomly selected from the original phage peptide library) and PBS were used as control groups. Binding was showed as a ratio of cancer/normal cell binding. Data in the first column of affinity values were reproduced from Fig. 1 in the reference and expressed as mean ± the standard deviation of three replicates. The values for IRP and PBS are 1.25 ± 0.10 and 1.05 ± 0.30, respectively. In addition, cell immunofluorescence assay was performed. Fluorescence images were observed using laser scanning confocal microscope (LSCM, Leica, Germany). Fluorescence signal intensity (IOD/Area) in the second column of affinity values was reproduced from Fig. 2B in the reference. Fluorescence signal intensity for IRP and PBS controls is 3.66155 ± 0.21256 and 2.04334, respectively. NT denotes not tested.

Brief Description:

Phages were firstly incubated with HEK293 cells. After incubation, the supernatant containing unbound phages was incubated with the blocked Eca-109 cells.

BiopanningDataSet Comments:

The 60 randomly selected phage clones were tested using cellular enzyme-linked immunosorbent assay (ELISA), and 41 phage clones were identified as positive clones with the over 2.10 ratio of absorbance higher than other clones, IRP and PBS controls. From the sequencing results of the positive clones, 14 peptide sequences were obtained and LLQLTTTKRPIT (ESCP9) consensus sequence was identified as the peptide with best affinity to ESCC cells via competitive inhibition, fluorescence microscopy, and flow cytometry. The results indicate that the peptide ESCP9 can bind to ESCC cells specifically and sensitively, and it is a potential candidate to be developed as an useful molecule to the imaging detection and targeting therapy for ESCC.

Curator:

Curation Date:

0000-00-00

First |

Prev |

2866

| Next

| Last