BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3507

Peptides:

Sequences

Affinity Values

YEHAMYRSAVLL
HDGTLLPRSSLH 
TIGVVRDVATQL
ISTVYTGLTEKD
DSLTMKRLVLPY
RNVELHDALRRT
NSSTPMLSDVLR
NRVSDHLVRAVS
HAPMPYANWPRG
HVKAAYPMVTMS

[1.67]
[2.16]
[2.06]
[1.31]
[1.58]
[1.38]
[1.70]
[6.71]
[1.78]
[3.42]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (common panning)

Sequencing Platform:

Illumina HiSeq

Sequencing Method:

Next-generation sequencing

Rounds of Panning:

Reference:

PMID: 31308251

Target Name:

Serum IgG of Systemic lupus erythematosus(SLE) patients

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:

Optical density was read at 450 nm using a Behring EL311 ELISA microplate reader (Dade Behring Marburg Gmbh, Berlin, Germany). A typical four-parameter logistic nonlinear regression model was used for standard curve fitting for the ELISA, which was further used to estimate sample content (units/ml) from the absorbance measurement data.

Brief Description:

The concentration of IgG from the samples were measured with the commercial Human IgG ELISA Quantitation kits (eBioscience, Massachusetts) according to the instructions. Besides the serum samples, we also added 1 μg of an anti-6×His antibody (Millipore, California) and 10 μl PBS buffer to a 100 μl immunoprecipitation mix as positive control and negative control, respectively. About 1 × 109 M13-phage particles were added to 100 μl immunoprecipitation mix in a 96-well PCR plate. The plate was then carefully sealed with adhesive optical tape (Applied Biosystems, California) and rotated overnight at 4 °C. 10 μl of protein G Dynabeads (Invitrogen, Massachusetts) were then added to each well. The resealed plate was placed back on a rotator for 4 h at 4 °C. The beads were washed six times in 200 μl wash buffer (150 mm NaCl, 50 mm Tris-HCl, 0.1% NP-40 (pH 7.5)) by pipetting up and down eight times per wash. After removal of the wash buffer, the beads were resuspended in 30 μl ddH2O and heated at 95 °C for 10 min.

BiopanningDataSet Comments:

To discover biomarkers for Systemic lupus erythematosus(SLE),a phage displayed random peptide library (Ph.D. 12) and deep sequencing were applied to screen specific autoantibodies in a total of 100 serum samples from 50 SLE patients and 50 healthy controls. Then, four peptides (YEHAMYRSAVLL, HDGTLLPRSSLH,TIGVVRDVATQL, and ISTVYTGLTEKD) were discovered with high diagnostic power to differentiate Systemic lupus erythematosus(SLE) patients from healthy controls. Among them, two peptides, i.e. YEHAMYRSAVLL and HDGTLLPRSSLH, were confirmed between SLE with other autoimmune patients.

Curator:

Chunchao Pu

Curation Date:

2020-12-29

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