BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3558

Peptides:

Sequences
WEYDRYRGWHIG(2) RWPPHFEWHFDD(1)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (common panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 32176482

Target Name:

Anti-F1 capsular antigen monoclonal antibody (YPF19) and human sera pool IgG of Alzheimer’s disease (AD) patients

Template Name:

F1 capsule antigen

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

pVIII-12aa phage display library (X12)

Affinity Measurement Method:

ELISA

Affinity Measurement Description:

Immunoscreening:positive spots on the immunoblots were detected using the Stable DAB chromogen system (Life Technologies, Monza, Italy). ELISA:the ELISA signal was measured at 450 nm using Labsystem Multiskan Bichromatic.

Brief Description:

Briefly, in the first round of selection, 500 μL of DYN−mAb YPF19 were incubated with 100 μL of each of the four phage libraries with a titer of e12 for 3−4 h at room temperature under mild stirring. The beads were washed 3 times in PBS−0.05% Tween 20 and separated with a magnetic device for 1−2 min to eliminate supernatants containing phage that did not bind the target present on the functionalized beads. Selected phage clones were eluted from antibodies with 500 μL of eluting buffer, 0.2 M of glycine-HCl (pH 2.2) + 0.1% BSA, neutralized immediately with 1 M Tris−HCl, pH 9.6. The enriched phage pools were amplified by infecting TG1 E. coli, purified twice by PEG precipitation, titrated, and used as the input for further panning. Biopanning affinity selection was repeated in the second round against DYN−IgG-AD, then in the third round against DYN−mAb YPF19 again, and finally a fourth selection round was carried out as the second one.

BiopanningDataSet Comments:

Peptide 12III1(RWPPHFEWHFDD), was found to be able to prevent in vitro Aβ1-42-induced cytotoxicity in SH-SY5Y cells, as well as to promote disaggregation of preformed fibrils, to a greater extent with respect to wild-type phage (pC89). IgG levels detected by 12III1 provided a significant level of discrimination between diseased and nondemented subjects, as well as a good correlation with the state progression of the disease. These results give significant impact in AD state and stage diagnosis, paving the way for the development not only for an innovative blood diagnostic assay for AD precise diagnosis, progressive clinical assessment, and screening but also for new effective treatments.

Curator:

Chunchao Pu

Curation Date:

2020-12-29

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