BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3472

Peptides:

Sequences

Affinity Values

CTGTPARQC(4/47)
CKNSMFATC(4/47)
CTNKHSPKC(3/47)
CSPKNILHC(3/47)
CFPSPTRTC(2/47)
CNAGTLGRC(2/47)
CNKEFASQC(2/47)
CNSANNRIC(2/47)
CSTQSTTSC(2/47)
CAPPGKSEC(1/47)
CAPKNILHC(1/47)
CAPSSSATC(1/47)
CESKTPKNC(1/47)
CFDHHTNSC(1/47)
CGTRETLSC(1/47)
CHAALNRSC(1/47)
CHHSNQRQC(1/47)
CHRPDTRSC(1/47)
CNRESPHLC(1/47)
CNSRSHAIC(1/47)
CPMPSTSYC(1/47)
CPVTSRSDC(1/47)
CRAWNEAPC(1/47)
CSDRGLPSC(1/47)
CSSKSDHSC(1/47)
CTAYPAKAC(1/47)
CTGAPARWC(1/47)
CTKTGLHIC(1/47)
CTSTAPLKC(1/47)
CVTSPFHNC(1/47)
CYSPRGGSC(1/47)
CYTNPDNVC(1/47)

[15.66 ± 2.82]
[10.56 ± 1.09]
[10.61 ± 0.73]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning, in vivo)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 30788426

Target Name:

Human prostate cancer cell line LNCaP

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:

Binding assay

Affinity Measurement Description:

LN1 (CTGTPARQC), LN2 (CKNSMFATC), and LN3 (CTNKHSPKC) peptides modified with a biotin molecule were incubated with LNCaP cells for 24 h at a concentration of 1 mg/mL and the binding affinities of the respective peptides were compared using fluorescence with Avidin D-conjugated TRITC (biotinylated peptides are stained red). The fluorescence intensity of each target peptide bound to LNCaP cells (n = 6 per group) was reporduced from Figure 2C and shown.

Brief Description:

The phage library was injected into C57BL/6 mice through the tail vein at a titer of 1.0e9 plaque-forming units (PFU)/mL in 100 mL Tris-buffered saline (50 mM Tris-HCl [pH7.5]). Five minutes after the injection, blood was recovered from the heart in order to remove non-specifically bound phages. The collected blood was dissolved in DMEM (Thermo Fisher Scientific) containing protease inhibitor (Sigma-Aldrich, St Louis, MO, USA). The solution was introduced into Escherichia coli ER2738 (New England Biolabs), and the number of phages in the blood was titrated by plaque-forming units. After amplification to 1.0e11 PFU, the phages were injected into new C57BL/6 mice. After three rounds of in vivo panning with mice, the phages were injected into the femoral vein of a 15-year-old female cynomolgus monkey. Five minutes after the injection, blood was taken from the femoral vein and dissolved in DMEM with protease inhibitor. The solution was centrifuged to collect the serum, from which phages were amplified to 1.0e11 PFU. These phages were injected into the tail vein of a C.B-17 SCID mouse with tumor xenografts (human prostate cancer cell line LNCaP (RCB2144)), and xenografted prostate cancer tissue was isolated after trans-cardiac removal of blood. Phages were recovered from the homogenized tissue and amplified as described above. Three additional cycles of in vivo phage panning were performed. After four cycles of in vivo panning in C.B-17 SCID mice, phage DNA was isolated from selected phage plaques with high affinity to prostate cancer tissue.

BiopanningDataSet Comments:

LN1 (CTGTPARQC), LN2 (CKNSMFATC), and LN3 (CTNKHSPKC) were synthesized and evaluated for binding and biological activity. LN1 showed the highest avidity for LNCaP prostate cancer cells in vitro and was thus administered to tumor-bearing mice to evaluate in vivo binding. Strikingly, LN1 specifically bound to the tumor tissue and exhibited very low reactivity with normal liver and kidney tissues. To demonstrate that LN1 could specifically deliver drugs to prostate cancer tissue, a therapeutic peptide, LN1-KLA (C-TGTPARQ-CGGG-D[KLAKLAK]2), was prepared and used to treat LNCaP cells in vitro and was also administered to tumor-bearing mice. The therapeutic peptide significantly suppressed growth of the cells both in vitro and in vivo.

Curator:

Bifang He

Curation Date:

2019-08-17

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