BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3007

Peptides:

Sequences

CAKATCPAC(26)
CSWQIGGNC(20)
CTVRTSADC(12)
CIGNSNTLC(10)
CIKVGKLQC(2)
CNWGDRILC(2)
CTNANHYFC(2)
CGWSGSLVC(1)
CLYANSPFC(1)
CSISSLTDC(1)
CNWGDRIRC(1)
CNWGDRIEC(1)
CRSANIFTC(1)
CFNMFSRFC(1)
CISAWPTRC(1)
CSISSLTHC(1)
CFLNPTTTC(1)
CKNLTRLAC(1)
CMARYMSAC(1)
CIKVGKVQC(1)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (in vivo)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 25366491

Target Name:

Human adenocarcinoma lung cancer cell line A549

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-C7C phage display library (CX7C)

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

For in vivo biopanning, a mouse bearing A549-derived xenograft tumor was injected with 1.0e9 pfu of the random library in TBS buffer 200 μl via the tail vein. After 15 min of biocirculation, the mouse was anesthetized by i.p. injection of zoletil (40 mg/kg) plus rompun (10 mg/kg) mixture and perfused with 50 mL of Tris-buffered saline (TBS; 50 mM Tris–HCl (pH 7.6), 150 mM NaCl) to remove non-specific and unbound phages. In subsequent rounds of biopanning to fourth rounds, different concentrations of detergent in the TBS buffer were used to wash out effectively non-specific bound phages. The increasing concentrations of Tween-20 were as follows: 0.1 % for second round, 0.3 % for third round and 0.5 % for fourth round in the TBS buffer. Then tumor tissues were resected from the mouse and homogenized in 1 mL of cold TBS containing 1× protease inhibitor cocktail (Tech and Innovation Co., Ltd, Seoul, Korea) on ice. Bound phages were subsequently eluted from the homogenate by 0.2 M glycine–HCl (pH 2.2) and the mixture was neutralized with 1 M Tris–HCl (pH 9.1).

BiopanningDataSet Comments:

Novel peptides were categorized into four groups according to a sequence-homology phylogenicity, and in vivo tumor-targeting capacity of these peptides was validated by whole body imaging with Cy5.5-labeled phages in various cancer types. The result revealed that novel peptides accumulated only in adenocarcinoma lung cancer cell-derived xenograft tissue. For further confirmation of the specific targeting ability, in vitro cell-binding assay and immunohistochemistry in vivo tumor tissue were performed with a selected peptide (G9, LYANSPF). The peptide was found to bind intensely to lung cancer cells both in vitro and in vivo, which was efficiently compromised with unlabeled phages in an in vitro competition assay. In conclusion, the peptides specifically targeting human lung cancer were discovered in this study, which is warranted to provide substantive feasibilities for drug delivery and imaging in terms of a novel targeted therapeutics and diagnostics.

Curator:

Curation Date:

0000-00-00

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