BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3445

Peptides:

Sequences
GLVSSQLTQFVIPAHCD(2)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (competitive panning)

Sequencing Platform:

Illumina MiSeq

Sequencing Method:

Next generation sequencing

Rounds of Panning:

Reference:

PMID: 30267550

Target Name:

Serum 13002 from patients sensitized to soy bean

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

ENTE-1 phage display library (GX3[C/S]X12)

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round.

BiopanningDataSet Comments:

Curator:

Bifang He

Curation Date:

2019-05-28

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