BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 1215

Peptides:

Sequences	Affinity Values
KSKRGHLLL HHRGAAPNS PIRVPGFLG PRLKHVQSV SQLRTRLKG PPRVPTTLR IPPALSISQ FRRTVNNKL RLNPHHEPM ATFTKDAAT	[2.870] [2.973] [0.110] [0.227] [NT] [NT] [NT] [NT] [NT] [NT]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (common panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 10921846

Target Name:

IgG fraction of anti-EHV-1 serum

Template Name:

Equine herpesvirus type 1

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

pVIII-9aa phage display library (X9)

Affinity Measurement Method:

ELISA

Affinity Measurement Description:

ELISA measurements of the binding to sera from EHV-1 infected gnotobiotic foals expressed as A490. The samples were taken in duplicate and the results are expressed as an average value. NT denotes not tested.

Brief Description:

Briefly phage (3e9-9e10 plaque forming units, pfu) in 100 ml PBSTG (phosphate buffered saline containing 0.05% Tween 20 and 0.5% goat sera, Seralabs) were incubated with or without biotinylated equine IgG for 5h at 37°C. An equal volume of M-280 streptavidin-coated magnetic beads (Dynal), washed three times with PBSTG, was added, mixed and stored at room temperature for 5 min.

BiopanningDataSet Comments:

The sequences of the amino acids displayed were aligned with protein sequences of EHV-1, thereby identifying a number of potential antibody binding regions. Presumptive epitopes were identified within the proteins encoded by genes 7 (DNA helicase:primase complex protein), 11 (tegument protein), 16 (glycoprotein C), 41 (integral membrane protein), 70 (glycoprotein G), 71 (envelope glycoprotein gp300), and 74 (glycoprotein E). Two groups of sequences, which aligned with either glycoprotein C (gC) or glycoprotein E (gE), identified type-specific epitopes which could be used to distinguish between sera from horses infected with either EHV-1 or EHV-4 in an ELISA using either the phage displaying the peptide or synthetic peptides as antigen. The gC epitope had been previously identified as an immunogenic region by conventional monoclonal antibody screening whereas the gE antibody binding region had not been previously identified.

Curator:

Curation Date:

0000-00-00

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