BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3548

Peptides:

Sequences

EIAYPARYANTY
IPWTQHMAMSPM
QNKLWDTPSNPW
TALGHQPLMRNT
SGGMPTARMSHQ
APWHNSWSEERT
ESGLWYSIDMKP
YLDEFAWYRFTH
WPRPYYGDWFQT
YPPPDSHSERVE
SMQGKAYGGTVM
WPRPYYGEGFQT
HPLTWNLRSSPA
ADWYHWRSHSSS
VVSPDMNLLLTN
TSLDGRISYHNR
VHWDFRQWWQPS
SQWETSQMIQKM
LPLTRGYVGDQY
FSPHADWVVVSG
GFAVGARDSLMF

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

ABI PRISM 377 sequencer from the Phage Display Peptide Library kit

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 30789148

Target Name:

Sera from 30 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:

Affinity Measurement Description:

Brief Description:

Serum pools were obtained from 30 HCs and 30 SLE patients. Microtitre wells were coated overnight at 4°C with 150 μl sera from healthy controls diluted with NaHCO3. Plates were blocked with 3% non-fat milk for 2 h at 37°C, and subsequently washed six times with phosphate-buffered saline with Tween 20 (0.05% Tween 20). A 100-μl diluted random 12-peptide phage display library with a titre of 2.0e11 pfu/ml was added to the coated plates. After incubation for 1 h at room temperature, unbound phages were collected and 100μl was added per well to SLE serumcoated plates. Following incubation for 1 h at room temperature, bound phages were eluted with 100 μl 0.2 mol/l glycine–HCl (pH 2.2) and neutralised with 1 mol/l Tris-HCl (pH 9.1). Eluted phages were amplified in a host strain and purified by precipitation for about 4 h using one-sixth volume of polyethylene glycol/NaCl. Another two rounds of SLE-serum affinity selection were carried out using the same process.

BiopanningDataSet Comments:

After a negative selection step with serum from healthy controls (HCs), a phage library of 12 peptides was used for three rounds of screening with sera from 30 SLE patients. After four rounds of biopanning, 21 positive peptides were sequenced. We produced 37-feature arrays containing 16 recombinant citrullinated peptides. The microarrays were tested with an independent validation set of serum samples from 50 HCs, 60 SLE patients, and 60 rheumatoid arthritis (RA) patients.

Curator:

Chunchao Pu

Curation Date:

2020-12-29

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