BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3228

Peptides:

Sequences	Affinity Values
CAVECPSTLDKH LSSNPKLYVGLE	[5.05, 4.35] [5.32, 4.42]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 28434129

Target Name:

Human IgG anti-HEV positive serum

Template Name:

Capsid protein

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:

The sandwich phage ELISA was used to test mimotope reactivity against anti-HEV positive sera S51 and S56, respectively. The ratio of clone OD absorbance and wt (wild-type phage M13KE) OD absorbance at 450 nm was calculated. The first column of the affinity values is the ratio with S51, the second column is the ratio with S56.

Brief Description:

The positive selection was carried out with protein G magnetic micro beads (Miltenyi Biotec) labelled with human IgG anti-hepatitis E virus (HEV) positive serum. The eluted bound phage was negatively selected with beads labelled with human IgG anti-HEV negative serum. Unbound phage from negative selection was amplified in E. coli ER2738 by a high-throughput method [38] and subjected to one more round of biopanning with different positive and negative sera.

BiopanningDataSet Comments:

Four mimotopes did not have homology with the primary sequence of HEV ORF2 capsid but competed effectively with a commercial HEV antigen for binding to anti-HEV reference serum. When the reactivity profile of each mimotope was compared with Wantai HEV-IgG ELISA, the most sensitive HEV immunoassay, mimotopes showed 95.2–100% sensitivity while the specificity ranged from 81.5 to 95.8%. PepSurf algorithm was used to map affinity-selected peptides onto the ORF2 crystal structure of HEV genotype 3, which predicted that these four mimototopes are clustered in the P domain of ORF2 capsid, near conformational epitopes of anti-HEV neutralising monoclonal antibodies. These HEV mimotopes may have potential applications in the design of structural vaccines and the development of new diagnostic tests.

Curator:

Bifang He

Curation Date:

2017-10-24

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