In Vivo Human Scfv Phage Display Assisted Identification Of Galectin-3 As A New Biomarker For Atherosclerosis

2019 ◽  
Vol 287 ◽  
pp. e166
M.J. Jacobin-Valat ◽  
A. Hemadou ◽  
A. Fontayne ◽  
C. Lorenzato ◽  
J. Laroche-Traineau ◽  
2021 ◽  
Yeon-Sook Choi ◽  
Myung Ji kim ◽  
Eun A Choi ◽  
Sinae Kim ◽  
Eun Ji Lee ◽  

Abstract Background One of the major challenges in pancreatic ductal adenocarcinoma (PDAC) management is a local or distant metastasis and limited targeted therapeutics to prevent the process. We aimed to identify a druggable target by screening abnormally secreted protein from PDAC and explore its therapeutic intervention. Methods A LC-MS/MS-based proteomics was carried out for TIF (Tumor Interstitial Fluids) obtained from patient derived xenograft (PDX) models of PDAC. To develop a blocking antibody for selected target protein, antibody phage-display technology was used. Results The proteomic screening of PDAC secretome identified Galectin-3 binding protein (Gal-3BP hereafter) as a top candidate. The Gal-3BP is highly expressed and secreted in PDAC tumors and primary cells. Subsequent functional tests by stable knockdown revealed the Gal-3BP is required for PDAC cell proliferation, migration and invasion. In addition, the depletion of Gal-3BP significantly abrogated in vivo tumor formation and metastasis of pancreatic cancer cells. Mechanistically, we found Gal-3BP enhances the galectin-3 mediated EGFR signaling, leading to the activation of cMyc and its target genes related to EMT. To examine the clinical usability of these findings, we screened a Gal-3BP-immunized chicken antibody library using phage display technique. The two isolated blocking antibody clones against Gal-3BP profoundly inhibited the metastasis of PDAC cells in vivo. Conclusions Altogether, our data demonstrates the Gal-3BP is an important therapeutic target in PDAC and proposed its blockade by antibody as a novel, therapeutic option for the inhibition of PDAC metastasis.

2018 ◽  
Vol 8 (1) ◽  
Audrey Hemadou ◽  
Jeanny Laroche-Traineau ◽  
Ségolène Antoine ◽  
Philippe Mondon ◽  
Alexandre Fontayne ◽  

2020 ◽  
Lei Peng ◽  
Chun Li ◽  
Tong Wu ◽  
Haiyan Zhang ◽  
Han Chen ◽  

Abstract BackgroundMethicillin-resistant Staphylococcus aureus (MRSA) is resistant to almost all β-lactam antibiotics. Hence, new ways to control MRSA infection, such as antibacterial antibodies, need to be explored. α-Hemolysin is the most important virulence factor widely expressed in S. aureus. This study aimed to develop a new fully human antibody against α-hemolysin of S. aureus and explore its neutralizing effect. ResultsThe single-chain antibody fragments(scFvs)against S. aureus were screened from a fully human scFv library using phage display technology with purified α-hemolysin protein. The selected scFvs had good binding affinities to α-hemolysin and were highly specific to S. aureus. The IgG-like scFv-Fc inserted into the pcDNA3.1 or pMH3 vector was expressed in HEK293F suspension cells to extend the antibody's in vivo half-life and restore Fc function. The size of purified scFv-Fc was about 55 kDa. The functions of expressed scFv-Fcs against α-hemolysin were validated. The results of A549 cytotoxicity assays showed that scFv10-Fc and scFv555-Fc had better protective effects than other scFv-Fcs on A549 cells. The results of anti-rabbit erythrocyte lysis assay confirmed that scFv555-Fc had a significant neutralizing effect on toxins. The scFv555-Fc was used to construct the docking model of antigen–antibody complexes using Discovery Studio software. It predicted that the key binding sites of α-hemolysin were TYR28, LYS37, PHE39, ARG56, and LYS58, which might be the key toxic sites of α-hemolysin.ConclusionsA novel fully human scFv-Fc antibody neutralizing the α-hemolysin toxin of S. aureus was successfully developed. The findings might provide a new theoretical basis and treatment method for preventing MRSA infection.

2002 ◽  
Vol 1591 (1-3) ◽  
pp. 87-97 ◽  
Ingrid N Michon ◽  
Arnaud D Hauer ◽  
Jan H von der Thüsen ◽  
Tom J.M Molenaar ◽  
Theo J.C van Berkel ◽  

2007 ◽  
Vol 15 (9) ◽  
pp. 1647-1654 ◽  
Laura Denby ◽  
Lorraine M Work ◽  
Dan J Von Seggern ◽  
Eugene Wu ◽  
John H McVey ◽  

1996 ◽  
Vol 14 (4) ◽  
pp. 429-431 ◽  
Anna M. Wu

2006 ◽  
Vol 4 (12) ◽  
pp. 51
B.-H. Lee ◽  
H.-Y. Hong ◽  
S.-J. Oh ◽  
E.-J. Lee ◽  
K. Wan ◽  

2006 ◽  
Vol 5 (2) ◽  
pp. 39
E.S. Yoo ◽  
B.H. Lee ◽  
T.G. Kwon ◽  
S.K. Chung ◽  
B.W. Kim ◽  

2004 ◽  
Vol 32 (3) ◽  
pp. 397-402 ◽  
E. Ruoslahti

In vivo screening of phage-displayed peptide libraries has revealed extensive molecular differences in the blood vessels of individual normal tissues. Pathological lesions also put their signature on the vasculature; in tumours, both blood and lymphatic vessels differ from normal vessels. The changes that characterize tumour blood vessels include selective expression of certain integrins. Peptides isolated by in vivo phage display for homing to tumours have been shown to be useful in directing therapeutic agents to experimental tumours. The targeting can enhance the efficacy of the therapy while reducing side effects. Phage screening has also revealed lung-specific vascular markers that promote tumour metastasis to the lungs by mediating specific adherence of tumour cells to the lung vasculature. These phage-screening studies have revealed a previously unsuspected degree of vascular specialization and provide potentially useful guidance devices for targeted therapies.

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