scholarly journals High-Throughput Method for Ranking the Affinity of Peptide Ligands Selected from Phage Display Libraries

2008 ◽  
Vol 19 (5) ◽  
pp. 993-1000 ◽  
Author(s):  
A. González-Techera ◽  
M. Umpiérrez-Failache ◽  
S. Cardozo ◽  
G. Obal ◽  
O. Pritsch ◽  
...  
Keyword(s):  
Phage Display ◽  
High Throughput ◽  
Peptide Ligands ◽  
Phage Display Libraries ◽  
High Throughput Method

Scanning whole cells with phage-display libraries: Identification of peptide ligands that modulate cell function

1994 ◽  
Vol 33 (2) ◽  
pp. 64-70 ◽  
Author(s):  
Susan Fong ◽  
Laura V. Doyle ◽  
James J. Devlin ◽  
Michael V. Doyle
Keyword(s):  
Phage Display ◽  
Cell Function ◽  
Peptide Ligands ◽  
Whole Cells ◽  
Phage Display Libraries

scholarly journals Rescue and In Situ Selection and Evaluation (RISE): A Method for High-Throughput Panning of Phage Display Libraries

2005 ◽  
Vol 10 (2) ◽  
pp. 108-117 ◽  
Author(s):  
Thomas Vanhercke ◽  
Christophe Ampe ◽  
Luc Tirry ◽  
Peter Denolf
Keyword(s):  
Phage Display ◽  
High Throughput ◽  
High Throughput Screening ◽  
Enzyme Linked Immunosorbent Assay ◽  
Selective Enrichment ◽  
Phage Display Libraries ◽  
Viral Particles ◽  
Phage Libraries ◽  
Rapid Characterization

Phage display has proven to be an invaluable instrument in the search for proteins and peptides with optimized or novel functions. The amplification and selection of phage libraries typically involve several operations and handling large bacterial cultures during each round. Purification of the assembled phage particles after rescue adds to the labor and time demand. The authors therefore devised a method, termed rescue and in situ selection and evaluation (RISE), which combines all steps from rescue to binding in a single microwell. To test this concept, wells were precoated with different antibodies, which allowed newly formed phage particles to be captured directly in situ during overnight rescue. Following 6 washing steps, the retained phages could be easily detected in an enzyme-linked immunosorbent assay (ELISA), thus eliminating the need for purification or concentration of the viral particles. As a consequence, RISE enables a rapid characterization of phage-displayed proteins. In addition, this method allowed for the selective enrichment of phages displaying a hemagglutinin (HA) epitope tag, spiked in a 104-fold excess of wild-type background. Because the combination of phage rescue, selection, or evaluation in a single microwell is amenable to automation, RISE may boost the high-throughput screening of smaller sized phage display libraries.

scholarly journals One-Step Recovery of scFv Clones from High-Throughput Sequencing-Based Screening of Phage Display Libraries Challenged to Cells Expressing Native Claudin-1

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Emanuele Sasso ◽  
Rolando Paciello ◽  
Francesco D’Auria ◽  
Gennaro Riccio ◽  
Guendalina Froechlich ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Virus Infection ◽  
Phage Display ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Hepatitis C Virus Infection ◽  
Novel Approach ◽  
Phage Display Libraries ◽  
Junction Protein

Expanding the availability of monoclonal antibodies interfering with hepatitis C virus infection of hepatocytes is an active field of investigation within medical biotechnologies, to prevent graft reinfection in patients subjected to liver transplantation and to overcome resistances elicited by novel antiviral drugs. In this paper, we describe a complete pipeline for screening of phage display libraries of human scFvs against native Claudin-1, a tight-junction protein involved in hepatitis C virus infection, expressed on the cell surface of human hepatocytes. To this aim, we implemented a high-throughput sequencing approach for library screening, followed by a simple and effective strategy to recover active binder clones from enriched sublibraries. The recovered clones were successfully converted to active immunoglobulins, thus demonstrating the effectiveness of the whole procedure. This novel approach can guarantee rapid and cheap isolation of antibodies for virtually any native antigen involved in human diseases, for therapeutic and/or diagnostic applications.

scholarly journals Peptide phage display in biotechnology and biomedicine

2016 ◽  
Vol 62 (5) ◽  
pp. 481-495 ◽  
Author(s):  
G.A. Kuzmicheva ◽  
V.A. Belyavskaya
Keyword(s):  
Phage Display ◽  
Cancer Cells ◽  
Genetically Modify ◽  
Peptide Ligands ◽  
Nano Materials ◽  
Human Interactome ◽  
Phage Display Libraries ◽  
Wide Range ◽  
Diagnostic Values ◽  
Peptide Phage Display

To date peptide phage display is one of the most common combinatorial methods used for identifying specific peptide ligands. Phage display peptide libraries containing billions different clones successfully used for selection of ligands with high affinity and selectivity toward wide range of targets including individual proteins, bacteria, viruses, spores, different kind of cancer cells and variety of nonorganic targets (metals, alloys, semiconductors etc.) Success of using filamentous phage in phage display technologies relays on the robustness of phage particles and a possibility to genetically modify its DNA to construct new phage variants with novel properties. In this review we are discussing characteristics of the most known non-commercial peptide phage display libraries of different formats (landscape libraries in particular) and their successful applications in several fields of biotechnology and biomedicine: discovery of peptides with diagnostic values against different pathogens, discovery and using of peptides recognizing cancer cells, trends in using of phage display technologies in human interactome studies, application of phage display technologies in construction of novel nano materials

Identification of a porcine TLR2-targeting peptide ligands using a cell-based phage display combined with high-throughput sequencing

2020 ◽  
Author(s):  
Su-Bin An ◽  
Seo-Ho Oh ◽  
Jun-Yeong Lee ◽  
Kwang-Hwan Choi ◽  
Chang-Kyu Lee ◽  
...  
Keyword(s):  
Phage Display ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Oral Route ◽  
Peptide Ligands ◽  
Mucosal Vaccine ◽  
Cell Targeting ◽  
M Cell ◽  
Phage Display Technique

Abstract M cell targeting is one of the critical issues to develop efficient mucosal vaccine design. In this study, peptide ligands with high affinity to porcine TLR2, which is highly expressed in M cells and play an important role in mucosal immune responses in pigs, were identified through the cell-based phage display technique combined with high-throughput sequencing. A random phage-peptide library was applied to the porcine TLR2 overexpressing cell line and total 85, 557 unique peptide sequences were identified from approximately 9.0 × 107 reads after three rounds of both subtractive and non-subtractive biopanning via high-throughput sequencing. Among the unique sequences, three candidate peptide sequences, NAGHLSQ, VPSKPGL, and RANLDGQ, were selected based on their abundance in the third round of biopanning. Consequently, NAGHLSQ showed the highest affinity exclusively to porcine TLR2 compared with other candidates and its binding mechanism was inferred to be directly associated with ligand binding site of the TLR2 through the in vitro competitive analysis. The peptide identified in this research could be used in development of effective porcine mucosal vaccine as an M cell targeting moiety to enhance the transport of antigens into the Peyer's patch via oral route.

scholarly journals SORTCERY—A High–Throughput Method to Affinity Rank Peptide Ligands

2015 ◽  
Vol 427 (11) ◽  
pp. 2135-2150 ◽  
Author(s):  
Lothar “Luther” Reich ◽  
Sanjib Dutta ◽  
Amy E. Keating
Keyword(s):  
High Throughput ◽  
Peptide Ligands ◽  
High Throughput Method

Retooling phage display with electrohydrodynamic nanomixing and nanopore sequencing

Lab on a Chip ◽  
2019 ◽  
Vol 19 (24) ◽  
pp. 4083-4092 ◽  
Author(s):  
Lyndon J. Raftery ◽  
Christopher B. Howard ◽  
Yadveer S. Grewal ◽  
Ramanathan Vaidyanathan ◽  
Martina L. Jones ◽  
...  
Keyword(s):  
Phage Display ◽  
High Throughput ◽  
High Throughput Screening ◽  
Nanopore Sequencing ◽  
Phage Display Libraries ◽  
Target Binding

High throughput screening of phage display libraries for target binding molecules using electrohydrodynamic nanomixing and nanopore sequencing.

Identifying Peptide Ligands for Barley α-Amylase 1 Using Combinatorial Phage Display Libraries

1998 ◽  
Vol 46 (9) ◽  
pp. 3852-3857 ◽  
Author(s):  
D. W. S. Wong ◽  
G. H. Robertson ◽  
S. J. Tillin ◽  
C. Wong
Keyword(s):  
Phage Display ◽  
Peptide Ligands ◽  
Phage Display Libraries
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