BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 2440

Peptides:

Sequences	Affinity Values
SLDSTHTHAPWP(3) TLADTHTHRPWT(2) LALDTPSHRPWT(2) FWGVTPHGELRS(1)	[1.687 ± 0.075] [1.523 ± 0.117] [1.486 ± 0.078] [1.491 ± 0.103]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 24478253

Target Name:

Hepatocellular carcinoma cell line HepG2

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:

ELISA

Affinity Measurement Description:

Phage ELISA was performed to evaluate the affinity of eight phage clones to HepG2 cells. Plates were read at 450 nm. Phage clones displaying IRPs and PBS treatments were used as negative controls. Triplicate determinations were performed. Absorbance at 450 nm was reproduced from the graph and expressed as means ± S.D. The absorbances of phage clones displaying IRPs and PBS treatments were 0.031 and 0.025 ± 0.006, respectively.

Brief Description:

HepG2 cells and HEK293 cells were separately seeded into six-well. The phage library was incubated with HEK293 cells. After incubation, culture medium containing the unbound phages was transferred to HepG2 cell monolayer cultures and incubated. Four rounds of biopanning were performed.

BiopanningDataSet Comments:

In phage ELISA, the PC28 clone (displaying SLDSTHTHAPWP) appeared to bind most effectively to HepG2 cells than the other clones. Immunofluorescence assay further evaluated the affinity of the PC28 phage clone binding to HepG2 cells. Besides, the synthetic peptide HCSP4, SLDSTHTHAPWP, can compete with the binding of the PC28 phage clone to HepG2 cells in competitive inhibition assay, which indicated that the binding of phage PC28 to target cells was mediated by the peptide HCSP4. The binding specificity of the FITC-conjugated HCSP4 to HepG2 cells was further validated by flow cytometry analysis.

Curator:

Curation Date:

0000-00-00

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