Screening and identification of a renal carcinoma specific peptide from a phage display peptide library
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Screening and identification of a renal carcinoma specific peptide from a phage display peptide library

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Specific peptide ligands to cell surface receptors have been extensively used in tumor research and clinical applications. Phage display technology is a powerful tool for the isolation of cell-specific peptide ligands. To screen and identify novel markers for renal cell carcinoma, we evaluated a peptide that had been identified by phage display technology. Methods A renal carcinoma cell line A498 and a normal renal cell line HK-2 were used to carry out subtractive screening in vitro with a phage display peptide library. After three rounds of panning, there was an obvious enrichment for the phages specifically binding to the A498 cells, and the output/input ratio of phages increased about 100 fold. A group of peptides capable of binding specifically to the renal carcinoma cells were obtained, and the affinity of these peptides to the targeting cells and tissues was studied. Results Through a cell-based ELISA, immunocytochemical staining, immunohistochemical staining, and immunofluorescence, the Phage ZT-2 and synthetic peptide ZT-2 were shown to specifically bind to the tumor cell surfaces of A498 and incision specimens, but not to normal renal tissue samples. Conclusion A peptide ZT-2, which binds specifically to the renal carcinoma cell line A498 was selected from phage display peptide libraries. Therefore, it provides a potential tool for early diagnosis of renal carcinoma or targeted drug delivery in chemotherapy.

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Tu et al. Journal of Experimental & Clinical Cancer Research 2011, 30:105
http://www.jeccr.com/content/30/1/105
RESEARCH Open Access
Screening and identification of a renal carcinoma
specific peptide from a phage display peptide
library
1*† 1† 1 1 1 1 1Xiangan Tu , Jintao Zhuang , Wenwei Wang , Liang Zhao , Liangyun Zhao , Jiquan Zhao , Chunhua Deng ,
1 2*Shaopeng Qiu and Yuanyuan Zhang
Abstract
Background: Specific peptide ligands to cell surface receptors have been extensively used in tumor research and
clinical applications. Phage display technology is a powerful tool for the isolation of cell-specific peptide ligands. To
screen and identify novel markers for renal cell carcinoma, we evaluated a peptide that had been identified by
phage display technology.
Methods: A renal carcinoma cell line A498 and a normal renal cell line HK-2 were used to carry out subtractive
screening in vitro with a phage display peptide library. After three rounds of panning, there was an obvious
enrichment for the phages specifically binding to the A498 cells, and the output/input ratio of phages increased
about 100 fold. A group of peptides capable of binding specifically to the renal carcinoma cells were obtained,
and the affinity of these to the targeting cells and tissues was studied.
Results: Through a cell-based ELISA, immunocytochemical staining, immunohistochemical staining, and
immunofluorescence, the Phage ZT-2 and synthetic peptide ZT-2 were shown to specifically bind to the tumor cell
surfaces of A498 and incision specimens, but not to normal renal tissue samples.
Conclusion: A peptide ZT-2, which binds specifically to the renal carcinoma cell line A498 was selected from
phage display libraries. Therefore, it provides a potential tool for early diagnosis of renal carcinoma or
targeted drug delivery in chemotherapy.
Keywords: Renal cell carcinoma, Phage display, Peptide, Targeting
Introduction However, metastatic disease recurs in a third of these
Renal cell carcinoma (RCC) accounts for 3% of all adult patients. The patients with metastatic RCC have a poor
malignancies and is the most lethal urological cancer. It prognosis with a median survival time of 1 to 2 years [3].
accounted more than 57, 000 new cases and 13, 000 can- Detection of RCC in early stages helps increase the life
cer-related deaths in the United States in 2009[1]. In expectancy of the patient [4]. Two diagnosis methods,
hisChina around 23, 000 new patients with RCC are diag- topathology and image procedures (computed tomography
nosed each year, and the incidence is increasing rapidly scan, ultrasonography, or magnetic resonance imaging)
due to the aging population [2]. Approximately 60% of provide increase the early detection of the RCC.
Histopatients have clinically localized disease at presentation, pathologically, although several promising biomarkers
with the majority undergoing curative nephrectomy. such as Carbonic anhydrase IX, B7-H1 and P53 for RCC
have been under investigation, none currently have been
validated or are inroutine use [5,6]. Therefore, some novel* Correspondence: txabs9988@163.com; yzhang@wfubmc.edu
† Contributed equally molecular markers must be screened and identified for
1Department of Urology, The First Affiliated Hospital, Sun Yat-sen University,
improving early diagnosis and prognosis ofRCC.
Guangzhou 510700, Guangdong, PR China
2 Phage display is a molecular diversity technology thatWake Forest Institute for Regenerative Medicine, Wake Forest University
Health Sciences, Winston-Salem, NC, 27157, USA allows the presentation of large peptide and protein
Full list of author information is available at the end of the article
© 2011 Tu et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons
Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in
any medium, provided the original work is properly cited.Tu et al. Journal of Experimental & Clinical Cancer Research 2011, 30:105 Page 2 of 6
http://www.jeccr.com/content/30/1/105
libraries on the surface of filamentous phage. Phage dis- repertoire of 12-mer peptide sequences that express
ranplay libraries permit the selection of peptides and pro- dom twelve-amino-acid sequences. Extensively
sequenteins, including antibodies, with high affinity and cing the naive library has revealed a wide diversity of
specificity for all targets. An important distinctive mark sequences with no obvious positional biases.
+
of this technology is the direct link that exists between The E. coli host strain ER2738 (a robust F strain with
the experimental phenotype and its encapsulated geno- a rapid growth rate) (New England Biolabs) was used
type. Phage display technology is a powerful tool for the for M13 phage propagation. The A498 and HK-2 cells
selection of cell-specific peptide ligands at present [7]. were cultured in DMEM supplemented with penicillin,
Some laboratories have applied this technology to isolate streptomycin, and 10% fetal bovine serum. Cells were
peptide ligands with good affinity and specificity for a harvested when subconfluent, and the total number of
variety of cell types. The specific ligands isolated from cells was counted using a hemocytometer.
phage libraries can be used in diagnostic probe,
therapeutic target validation, and drug design and vaccine In Vitro Panning
development [8-10]. A498 cells were taken as the target cells, and HK-2 as
In the present study, we identified a specific novel the absorber cells for a whole-cell subtractive screening
peptide that bound to the cell surface of renal carci- from a phage display 12-peptide library. Cells were
culnoma cell line A498 generated in this laboratory by turedinDMEMwith10%FCSat37°Cinahumidified
using in vitro phage-displayed random peptide libraries. atmosphere containing 5% CO . HK-2 cells were washed2
Our results demonstrate that this biopanning strategy with PBS and kept in serum-free DMEM for 1 h before
can be used to identify tumor-specific targeting peptides. blocking with 3 mL blocking buffer (BF, PBS + 5% BSA)
11One of our selected peptides, ZT-2 was most effective in for 10 min at 37°C. Approximately 2 × 10 pfu phages
targeting cells and tissues, indicating its potential for use were added and mixed gently with the blocked HK-2 for
in early diagnosis and targeted therapy of RCC. 1 h at 37°C. Cells were then pelleted by centrifuging at
1000 rpm (80 g)for5min.HK-2andphagesboundto
Materials these cells were removed by centrifugation. Those
Renal carcinoma line A498 and a normal renal cell line phages in the supernatant were incubated with the
BFHK-2 were obtained from Medical Academy of China blocked A498 cells for 1 h at 37°C before cells were
pel(Beijing, PR China). Fetal calf serum (FCS) and Dulbecco’s leted again. After that, the pelleted cells were washed
modified eagle’s medium (DMEM) were purchased from twice with 0.1% TBST (50 mM Tris-HCl, pH 7.5, 150
Gibco(Invitrogen,Carlsbad,USA).PhageDNAsequen- mM NaCl, 0.1% Tween-20) to remove unbound phage
cing was performed by Shanghai Sangon Corp (Shanghai, particles. A498 cells and bound phages were both
incuPR China). Peptide ZT-2 (QQPPMHLMSYAG) and a bated with the E. coli host strain ER2738. Then, the
nonspecific control peptide (EAFSILQWPFAH) were phages were rescued by infection with bacteria while the
synthesized and labeled with fluorescein isothiocyanate cells died. The phage titer was subsequently evaluated
(FITC) by Shanghai Bioengineering Ltd. Mass analysis of by a blue plaque-forming assay on agar plates containing
the peptides was confirmed by a matrix-assisted laser des- tetracycline. Finally, a portion of purified phage
preparaorption/ionization time-of-flight mass spectrometry, and tion was used as the input phage for the next round of
all peptides were > 90% pure as determined by reverse- in vitro selection.
11phase HPLC. Peptide stock solutions were prepared in For each round of selection, more than 1.5 × 10 pfu
PBS (pH 7.4). Horseradish peroxidase-conjugated sheep of collected phages were used. The panning intensity
anti-rabbit antibody and rabbit anti-M13 bacteriophage was increased by prolonging the phage incubation
perantibody were purchased from Pharmacia (Peapack, NJ, iod with HK-2 for 1.25 h or 1.5 h, shortening the phage
USA). Trizol reagents were purchased from Gibco BRL incubation with A498 for 45 min and 30 min in the
sec(Gaithersburg, MD, USA) and the reverse transcriptase ond and third rounds individually, and increasing
washpolymerase chain reaction (RT-PCR)system kits werepur- ing with TBST for 4 times and 6 times in the second
chased from Promega (Madison, WI, USA). and third round individually.
ThePh.D.-12phagedisplaypeptidelibrarykit(New
England Biolabs, Beverly, MA, USA) was used to screen Sequence Analysis of Selected Phages and Peptide
specific peptides binding to A498 cells. The phage dis- Synthesis
play library contains random peptides constructed at the After three rounds of in vitro panning, 60 blue plaques
N terminus of the minor coat protein (cpIII) of the M13 were randomly selected and their sequences were
ana13
phage. The titer of the library is 2.3 × 10 pfu (plaque- lyzed with an ABI Automatic DNA Analyzer (Shanghai
forming units). The library contains a mixture of 3.1 × Sangon Corp). A primer used for sequencing was
5’9
10 individual clones, representing the entire obtainable CCC TCATAG TTAGCG TAACG-3’ (-96 gIIITu et al. Journal of Experimental & Clinical Cancer Research 2011, 30:105 Page 3 of 6
http://www.jeccr.com/content/30/1/105
sequencing primer, provided in the Ph.D.-12 Phage dis- of the First Affiliated Hospital of Sun Yat-Sen University
play peptide library kit). Homologous analysis and mul- (NO.2011-137), and oral or written informed consent
tiple sequence alignment were done using the BLAST was obtained from all subjects prior to enrollment in
and Clustal W programs to determine the groups of the study.
related peptides.
Peptide Synthesis and Labeling
Cell-Based ELISA with Phage The ZT-2 peptide (QQPPMHLMSYAG) translated from
A498 and HK-2 were cultured in DMEM with 10% FCS the selected M13 phage DNA sequence and nonspecific
at 37°C in a humidified atmosphere containing 5% CO , control peptide (EAFSILQWPFAH) were synthesized2
5and the cells were seeded into 96-well plates (1 × 10 and purified by Shanghai Bioengineering Ltd.
Fluorescells/well) overnight. Cells were then fixed on 96-well cein isothiocyanate (FITC)-conjugated peptides were
plates by 4% paraformaldehyde for 15 min at room tem- also produced by the same company.
perature until cells were attached to the plates.
Triplicate determinations were done at each data point. Peptide Competitive Inhibition Assay for Characterization
Selectivity was determined using a formula as follows of Specific Phage Clones
[11]: Selectivity = OD -OD /OD -OD . Here, The in vitro blue-plaque forming assay was performedM13 C1 S2 C2
OD and OD represent the OD values from the to observe the competitive inhibition effect of ZT-2 pep-M13 C1
selected phages and control phages binding to A498 tide with its phage counterparts (M13). A498 cells were
cells, respectively. OD and OD represent the OD cultured in a 12-well plate overnight and then preincu-S2 C2
values from the selected phage and control phage bind- bated with blocking buffer to block nonspecific binding
ing to the control (HK-2 cell line), respectively. at 4°C for 30 min. The synthetic peptide (0, 0.0001,
0.001, 0.01, 0.1, 1 or 10 μM) was diluted in PBS and
Immunocytochemical Staining and Immunohistochemical incubated with cells at 4°C for 1 h, and then incubated
11Staining of Phage M13 with 1 × 10 pfu of phage M13 at 4°C for 1 h. The
Before staining with phage M13 [12], the cells in the dif- bound phages were recovered and titered in ER2738
culferent groups (A498 and HK-2) were cultured on cover- ture. The phages binding to A498 cells were evaluated
slips and fixed with acetone at 4°C for 20 min. Then, by blue plaque-forming assay, and the rate of inhibition
11
about 1 × 10 pfu of phage M13 diluted in PBS were was calculated by the following formula: Rate of
inhibiadded onto the coverslips and incubated at 4°C over- tion = (number of blue plaques in A498 incubated with
night. Coverslips were then washed for five times with PBS - number of blue plaques in A498 with ZT-2
pepTBST. The coverslips were blocked by H O (3% in tide)/number of blue plaques in A498 incubated with2 2
PBS) at room temperature for 510 min. After being PBS × 100%. Nonspecific control phages (a synthetic
washed by PBS for 5 min at 37°C, the coverslips were peptide corresponding to an unrelated phage picked
incubated with normal sheep serum for 20 min at 37°C. randomly from the original phage peptide library) were
Subsequently, the coverslips were incubated overnight at used as negative controls.
4°C with a mouse anti-M13 phage antibody at a dilution
of 1:5000. The next day, the coverslips were rinsed for Immunofluorescence Microscopy and Image Analysis
three times (10 min for each rinse) in PBS and incu- microscopy was used to study the
bated with a secondary antibody for 1 h at room tem- affinity of synthetic peptide (ZT-2) binding to A498 and
perature. Afterward, the coverslips were rinsed three renal carcinoma. A498 and HK-2 were digested with
times (5 min for each rinse) in PBS. The bound anti- 0.25% trypsin and plated on coverslips overnight. Cells
body was visualized using DAB. The coverslips were were washed three times with PBS and fixed with
acetrinsed for three times (5 min for each rinse) using run- one at 4°C for 20 min before analysis. ZT-2 peptide
ning tap water before staining by hematoxylin and labeled with FITC was incubated with cells. PBS and
eosin. Finally, the coverslips were rinsed for 10 min with control peptides labeled with FITC were used as
negarunning tap water before dehydration and mounting. tive controls. After being washed for three times with
Frozen sections of human renal tissues with and with- PBS, the slips were observed using a fluorescence
out tumors were also prepared. The steps of immuno- microscope.
histochemical staining were similar to those for
immunocytochemical staining described above. Instead Results
of the selected phage clone M13, PBS and a nonspecific Specific Enrichment of A498 Cell-Bound Phages
control phage with same titers were used for negative Phages specifically bound to human A498 cells were
controls. The study protocol was reviewed and approved identified through three rounds of in vitro panning. In
by the Institutional Review Board and Ethic Committee each round, the bound phages were rescued andTu et al. Journal of Experimental & Clinical Cancer Research 2011, 30:105 Page 4 of 6
http://www.jeccr.com/content/30/1/105
amplified in E. coli for the following round of panning,
while the unbound phages were removed by washing
with TBST. After the third round of the in vitro
selection, the number of phages recovered from A498 cells
increased 100-fold (Table 1). However, the number of
phages recovered from HK-2 control cells decreased.
The output/input ratio of phages recovered after each
round of the panning was used to determine the phage
recovery efficiency. These results indicated an obvious
enrichment of phages specifically binding to A498 cells.
Figure 1 Evaluation by cell-ELISA of the binding selectivity of
twenty phage clones. The selectivity values of five higher phage
Verification of In Vitro Specific Binding by Cell-Based
clone (ZT-2, ZT-4, ZT-8, ZT-9, and ZT-16), calculated by the formula
ELISA mentioned in the text, were 3.15, 2.90, 2.95, 2.80, and 3.05,
A cellular ELISA was used to identify the affinities for respectively. Therefore, clone ZT-2 appeared to bind more
effectively than the other clones.the twenty selected phages binding to A498. To assess
selectivity, the affinities of each phage binding to A498
cells and to the control HK-2werecompared.These
unrelated phage clone or the normal renal tissue
secphage clones bound more effectively to A498 cells
comtions when bound with phage clone ZT-2 showed
negapared with PBS and HK-2 control groups. Furthermore,
tive staining. It is thus clear that the phage clone ZT-2
the ZT-2 clone appeared to bind most effectively to
was able to bind specifically to A498 cells (Figure 3).
A498 cells than the other clones (Figure 1). Therefore,
we further analyzed the phage M13 and its displaying
Competitive Inhibition Assay
peptide ZT-2.
A peptide-competitive inhibition assay was performed to
discover whether the synthetic peptide ZT-2 and the
Affinity of the Phage M13 to A498 Cells and Renal
corresponding phage clone competed for the same
carcinoma Tissues
To confirm the binding ability of the selected phage
toward target A498 cells, the phage clone M13 (clone
ZT-2) was isolated, amplified and purified for
immunochemical assay. The HK-2 cell line, composed of human
nontumor renal tissues, was included as a negative
control. The interaction of the M13 phage and target cells
(A498) was evaluated by immunocytochemical staining.
A498 cells bound by the phage M13 were stained brown
in contrast to the HK-2 cells. Negative results were also
obtained when A498 cells bound with unrelated phage
clone. However, A498 cells with phage clone
ZT2 were stained brown distinctively, demonstrating that
ZT-2 was able to bind specifically to A498 cells (Figure
2). Subsequently, immunohistochemical stain was
performed to observe the specific binding of the phage
clone ZT-2 toward human renal carcinoma tissues. The
cells in A498 tumor tissue sections when bound with
phage clone ZT-2 were stained green fluorescence
distinctively. When A498 tumor tissue sections bound by
Figure 2 Immunocytochemical staining of A498 and control
cells when bound with phage ZT-2. Cell-bound phages were
detected using anti-M13 monoclonal antibody, secondaryTable 1 Enrichment of phages for each round of
antibody, and ABC complex. The cells were stained withselection from phage displayed peptide library
diaminobenzidine (DAB). (A) shows control cell (B) shows
Rounds Selected Phage Eluted Phage Ratio
immunocytochemical staining of A498 cells when bound with
(input) (cpu) (output) (cpu) (output/input)
phages without exogenous sequences (wild-type phage) (C) shows
11 3 -8
1 1.5 × 10 1.5 × 10 1×10 immunocytochemical staining of A498 cells when bound with
12 5 -7
210 10 10 unrelated phage (D) shows immunocytochemical staining of A498
12 6 -6 cells when bound with phage ZT-2. Amplification × 200.310 10 10Tu et al. Journal of Experimental & Clinical Cancer Research 2011, 30:105 Page 5 of 6
http://www.jeccr.com/content/30/1/105
Figure 3 Immunohistochemical staining of renal carcinoma and nontumorous renal tissue sections when bound with ZT-2
peptidefluorescein isothiocyanate. To investigate if the free ZT-2 peptide maintained its binding affinity to renal carcinoma cells, we made a synthetic
peptide ZT-2 (QQPPMHLMSYAG) labeled with fluorescein isothiocyanate. (A) Immunohistochemical staining of renal carcinoma tissues when
bound with phage ZT-2-FITC. The specific binding sites on tumor cells fluoresced green (B) Immunohistochemical staining of nontumorous renal
tissues when bound with phage ZT-2 (C) a negative control section stained with random peptide-fluorescein isothiocyanate in renal carcinoma
tissues. Magnification × 200.
binding site. When the synthetic peptide ZT-2 was pre- often display certain cell surface antigens such as
incubated with A498 cells, phage ZT-2 binding to A498 tumor-associated antigens or tumor-specific antigens in
cells decreased in a dose-dependent manner. When the high quantity, which are different from the antigens on
peptide ZT-2 concentrations increased, the titer of normal tissues. To develop more biomarkers for the
phages recovered from A498 cells was decreased and diagnosis of RCC, we used peptide phage display
techthe inhibition was increased gradually. When the con- nology to identify potential molecular biomarkers of
centrations of peptide ZT-2 increased above 5 μM, the A498 carcinoma cells. After panning for three rounds,
inhibition reached a flat phase. The control peptide 20 clones were selected for further characterization.
(EAFSILQWPFAH) had no effect on the binding of the First, a cell-based ELISA assay was used to confirm the
phage ZT-2 to A498 cells (Figure 4). specific binding of the phage clones to A498 cells in
vitro. ZT-2 was the best candidate phage clone with the
Discussion highest specificity. Second, immunocytochemical and
Targeting specific ligand binding on specific tumor anti- immunohistochemical staining were performed to
congens is an efficient way to increase the selectivity of firm the selectivity of the phage ZT-2 to bind to A498
therapeutic targets in clinical oncology and helpful for cells. Third, the results of the competitive inhibitory
the early detection and therapy of RCC. Tumor cells assays suggest that the peptide displayed by the phage
M13-ZT-2, not other parts of this phage, can bind to
the renal carcinoma cell surface. Under the same
conditions, the normal renal cell line HK-2 did not show
significant fluorescence when stained with ZT-2
peptideFITC, which confirmed the targeting of ZT-2 to be
A498 cells.
Monoclonal antibodies have become the most rapidly
expanding class of drugs for treating kidney cancer, but
poor tumor penetration, bone marrow toxicity and high
immunogenicity of these antibodies have been limited in
clinical applications [13,14]. Compared with monoclonal
antibodies, peptide ligands, which have the advantages
of rapid tissue penetration, faster blood clearance, easy
incorporation into certain delivery vectors and low
immunogenicity are being pursued as targeting moietiesFigure 4 Competitive inhibition of binding of the phage ZT-2
to A498 cells by the synthetic peptide ZT-2 QQPPMHLMSYAG. for the selective delivery of radionuclides cytokines,
cheThe average inhibition rates at different concentrations of the mical drugs, or therapeuticgenestotumors[15].This
peptide are shown. When the concentration of the peptide ZT-2 effect may open up diagnostic procedures and
therapeureached more than 0.001 μM, a significant inhibition occurred.
tic options for the patient. Identification of the cancerTu et al. Journal of Experimental & Clinical Cancer Research 2011, 30:105 Page 6 of 6
http://www.jeccr.com/content/30/1/105
10. Shadidi M, Sioud M: Identification of novel carrier peptides for thecell receptors that binds the ZT-2 peptide would allow
specific delivery of therapeutics into cancer cells. FASEB J 2003,
further improvement of the peptide for potential clinical
17(2):256-258.
use. 11. Du B, Qian M, Zhou ZL, Wang P, Wang L, Zhang X, Wu M, Zhang P, Mei B:
In vitro panning of a targeting peptide to NCI-H1299 from a phageThese preliminary experiments provide evidence that
display peptide library. Biochem Biophys Res Comm 2006, 32(3):956-962.
the ZT-2 peptide may be specific to A498 and therefore
12. Yang XA, Dong XY, Qiao H, Wang YD, Peng JR, Li Y, Pang XW, Tian C,
it would be useful for diagnosis of renal carcinoma or Chen WF: Immunohistochemical analysis of the expression of
FATE/BJHCC-2 antigen in normal and malignant tissues. Lab Invest 2005,delivery of an antitumor therapeutic agent. Studies are
85(2):205-213.
continuing to identify the cellular receptors responsible
13. Davis ID, Liu Z, Saunders W, Lee FT, Spirkoska V, Hopkins W, Smyth FE,
for peptide binding and to apply the peptide to clinically Chong G, Papenfuss AT, Chappell B, Poon A, Saunder TH, Hoffman EW,
Old LJ, Scott AM: A pilot study of monoclonal antibody cG250 and lowrelevant samples.
dose subcutaneous IL-2 in patients with advanced renal cell carcinoma.
Cancer Immun 2007, 7:13.
14. Xu C, Lo A, Yammanuru A, Tallarico AS, Brady K, Murakami A, Barteneva N,
Acknowledgements
Zhu Q, Marasco WA: Unique biological properties of catalytic domain
This work was supported by National Natural Science Foundation of China
directed human anti-CAIX antibodies discovered through phage-display
(No.81172432), The Project Supported by Guangdong Natural Science
technology. PLoS One 2010, 5(3):e9625.
Foundation of the People’s Republic of China (No.9151802904000002),
15. Langer M, Beck-Sickinger AG: Peptides as carrier for tumor diagnosis and
Scientific and Technical Project of Guangdong Province of the People’s
treatment. Curr Med Chem Anticancer Agents 2001, 1(1):71-93.
Republic of China (2008B030301082), Doctoral Initiating Project, and Natural
Scientific Foundation of Guangdong Province of the People’s Republic of doi:10.1186/1756-9966-30-105
China (No.7301521) Cite this article as: Tu et al.: Screening and identification of a renal
carcinoma specific peptide from a phage display peptide library. Journal
Author details of Experimental & Clinical Cancer Research 2011 30:105.
1Department of Urology, The First Affiliated Hospital, Sun Yat-sen University,
2Guangzhou 510700, Guangdong, PR China. Wake Forest Institute for
Regenerative Medicine, Wake Forest University Health Sciences,
WinstonSalem, NC, 27157, USA.
Authors’ contributions
TXA and ZYY designed the study. ZJT performed the cell-based ELISA and
analyzed the data statistically. WWW performed immunocytochemical
staining. ZL performed immunohistochemical staining. ZLY and ZJQ
performed immunofluorescence microscopy and image analysis. DCH and
QSP performed data analysis. TXA wrote the main manuscript. ZYY looked
over the manuscript. All authors read and approved the final manuscript.
Competing interests
The authors declare that they have no competing interests.
Received: 30 May 2011 Accepted: 10 November 2011
Published: 10 November 2011
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