BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 1654

Peptides:

Sequences

Affinity Values

INTLLSEINSILLDIISLL(13)
INLLLMEINMILNEIITLL(3)
INTLLMEINNILLNIIDFL(2)
INTLLEEINIILNNIISFL(2)
INSLLKEINNILISIIDFL(2)
INSLLMEINTILNDIIMFL(1)
ISSLLEEINTILMDIIEYL(1)

[2.709 ± 0.168]
[3.409 ± 0.061]
[3.088 ± 0.161]
[3.079 ± 0.124]
[2.709 ± 0.113]
[2.983 ± 0.185]
[2.726 ± 0.079]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

mRNA display (competitive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 21136209

Target Name:

Interleukin-6, IL-6

Template Name:

Interleukin-6 receptor

Structure of Target-Template Complex:

1P9M

Structure of Target-Peptide Complex:

Not determined.

Library Name:

X(3)-L-I-X-E-I-X(2)-I-L-X(2)-I-X(3)-L mRNA display library

Affinity Measurement Method:

ELISA

Affinity Measurement Description:

In peptide ELISA, the absorbance at 490 nm was determined. Data were reproduced from the graph and expressed as the mean ± SD of triplicate determinations. The phage RNA(-) acted as negative control with A490 of 0.037. Beside, both RA07 (INTLLSEINSILLDIISLL) and CA11 (NQQLIEEIIQILHKIFEIL) peptides bound to GST-IL-6 in a dose-dependent manner and RA07 had higher affinity for GST-IL-6 (EC50=45.1 nM) than CA11 (EC50=142 nM). To determine whether CA11 and RA07 have inhibitory activity against IL-6, a KT-3 cell-based assay was performed. KT-3 cells are known to proliferate in response to IL-6. Results showed that RA07 suppressed KT-3 cell proliferation in a dose-dependent manner, but CA11 did not. The IC50 value of RA07 was 5.6 μM. Besides, the results of ELISA-based IL-6 inhibition assay suggested that RA07 inhibits gp130 from binding to the IL-6/IL-6R complex, but not IL-6R from binding to IL-6. The IC50 value of RA07 in this assay was 2.8 μM, which was almost the same as in the KT-3 cell-based assay (5.6 μM).

Brief Description:

After the beads were washed three times with selection buffer, 1-8 μM of IL-6 (Ajinomoto) was added and incubation continued another 2 h. The bound molecules were eluted with PreScission protease (GE healthcare).

BiopanningDataSet Comments:

The most frequently occurring sequence, RA07 (INTLLSEINSILLDIISLL), bound to IL-6 with 3 to 4-fold higher affinity than CA11. Furthermore, RA07 inhibited IL-6-dependent KT-3 cell proliferation in a dose-dependent manner. ELISAs revealed that RA07 could not inhibit IL-6 from binding to the IL-6 receptor (IL-6R), but could inhibit the IL-6/IL-6 complex binding to gp130.

Curator:

Curation Date:

0000-00-00

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