High-Throughput Platform for B-Cell Screening Based on Fluorescent Phage-Display Technology

2019 ◽  
Vol 167 (4) ◽  
pp. 446-451 ◽  
Author(s):  
Ya. A. Lomakin ◽  
A. N. Kaminskaya ◽  
A. V. Stepanov ◽  
A. A. Shmidt ◽  
A. G. Gabibov ◽  
...  
Keyword(s):  
Phage Display ◽  
B Cell ◽  
High Throughput ◽  
Phage Display Technology ◽  
Display Technology

Development and Application of Computational Methods in Phage Display Technology

2020 ◽  
Vol 26 (42) ◽  
pp. 7672-7693 ◽  
Author(s):  
Bifang He ◽  
Anthony Mackitz Dzisoo ◽  
Ratmir Derda ◽  
Jian Huang
Keyword(s):  
Phage Display ◽  
Computational Methods ◽  
Peptide Ligands ◽  
Next Generation ◽  
Phage Display Technology ◽  
Next Generation Sequencing Technology ◽  
Screening Experiments ◽  
Display Technology ◽  
Comprehensive Search ◽  
Generation Sequencing

Background: Phage display is a powerful and versatile technology for the identification of peptide ligands binding to multiple targets, which has been successfully employed in various fields, such as diagnostics and therapeutics, drug-delivery and material science. The integration of next generation sequencing technology with phage display makes this methodology more productive. With the widespread use of this technique and the fast accumulation of phage display data, databases for these data and computational methods have become an indispensable part in this community. This review aims to summarize and discuss recent progress in the development and application of computational methods in the field of phage display. Methods: We undertook a comprehensive search of bioinformatics resources and computational methods for phage display data via Google Scholar and PubMed. The methods and tools were further divided into different categories according to their uses. Results: We described seven special or relevant databases for phage display data, which provided an evidence-based source for phage display researchers to clean their biopanning results. These databases can identify and report possible target-unrelated peptides (TUPs), thereby excluding false-positive data from peptides obtained from phage display screening experiments. More than 20 computational methods for analyzing biopanning data were also reviewed. These methods were classified into computational methods for reporting TUPs, for predicting epitopes and for analyzing next generation phage display data. Conclusion: The current bioinformatics archives, methods and tools reviewed here have benefitted the biopanning community. To develop better or new computational tools, some promising directions are also discussed.

Identification of Small Molecule Binding Sites within Proteins Using Phage Display Technology

2001 ◽  
Vol 4 (7) ◽  
pp. 553-572 ◽  
Author(s):  
D. Rodi ◽  
G. Agoston ◽  
R. Manon ◽  
R. Lapcevich ◽  
S. Green ◽  
...  
Keyword(s):  
Phage Display ◽  
Small Molecule ◽  
Binding Sites ◽  
Phage Display Technology ◽  
Display Technology

scholarly journals High-Throughput Sequencing of Phage Display Libraries Reveals Parasitic Enrichment of Indel Mutants Caused by Amplification Bias

2021 ◽  
Vol 22 (11) ◽  
pp. 5513
Author(s):  
Sander Plessers ◽  
Vincent Van Deuren ◽  
Rob Lavigne ◽  
Johan Robben
Keyword(s):  
Amino Acid ◽  
Phage Display ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Target Selection ◽  
Acid Modification ◽  
Phage Display Technology ◽  
Amplification Bias ◽  
Large Deletions ◽  
Depth Analysis

The combination of phage display technology with high-throughput sequencing enables in-depth analysis of library diversity and selection-driven dynamics. We applied short-read sequencing of the mutagenized region on focused display libraries of two homologous nucleic acid modification eraser proteins—AlkB and FTO—biopanned against methylated DNA. This revealed enriched genotypes with small indels and concomitant doubtful amino acid motifs within the FTO library. Nanopore sequencing of the entire display vector showed additional enrichment of large deletions overlooked by region-specific sequencing, and further impacted the interpretation of the obtained amino acid motifs. We could attribute enrichment of these corrupted clones to amplification bias due to arduous FTO display slowing down host cell growth as well as phage production. This amplification bias appeared to be stronger than affinity-based target selection. Recommendations are provided for proper sequence analysis of phage display data, which can improve motive discovery in libraries of proteins that are difficult to display.

scholarly journals Uniqueness of RNA Coliphage Qβ Display System in Directed Evolutionary Biotechnology

Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 568
Author(s):  
Godwin W. Nchinda ◽  
Nadia Al-Atoom ◽  
Mamie T. Coats ◽  
Jacqueline M. Cameron ◽  
Alain Bopda Waffo
Keyword(s):  
Life Cycle ◽  
Coat Protein ◽  
Phage Display ◽  
Display System ◽  
Phage Display Technology ◽  
Library Size ◽  
Exterior Surface ◽  
Major Coat Protein ◽  
Display Technology ◽  
Minor Coat Protein

Phage display technology involves the surface genetic engineering of phages to expose desirable proteins or peptides whose gene sequences are packaged within phage genomes, thereby rendering direct linkage between genotype with phenotype feasible. This has resulted in phage display systems becoming invaluable components of directed evolutionary biotechnology. The M13 is a DNA phage display system which dominates this technology and usually involves selected proteins or peptides being displayed through surface engineering of its minor coat proteins. The displayed protein or peptide’s functionality is often highly reduced due to harsh treatment of M13 variants. Recently, we developed a novel phage display system using the coliphage Qβ as a nano-biotechnology platform. The coliphage Qβ is an RNA phage belonging to the family of Leviviridae, a long investigated virus. Qβ phages exist as a quasispecies and possess features making them comparatively more suitable and unique for directed evolutionary biotechnology. As a quasispecies, Qβ benefits from the promiscuity of its RNA dependent RNA polymerase replicase, which lacks proofreading activity, and thereby permits rapid variant generation, mutation, and adaptation. The minor coat protein of Qβ is the readthrough protein, A1. It shares the same initiation codon with the major coat protein and is produced each time the ribosome translates the UGA stop codon of the major coat protein with the of misincorporation of tryptophan. This misincorporation occurs at a low level (1/15). Per convention and definition, A1 is the target for display technology, as this minor coat protein does not play a role in initiating the life cycle of Qβ phage like the pIII of M13. The maturation protein A2 of Qβ initiates the life cycle by binding to the pilus of the F+ host bacteria. The extension of the A1 protein with a foreign peptide probe recognizes and binds to the target freely, while the A2 initiates the infection. This avoids any disturbance of the complex and the necessity for acidic elution and neutralization prior to infection. The combined use of both the A1 and A2 proteins of Qβ in this display system allows for novel bio-panning, in vitro maturation, and evolution. Additionally, methods for large library size construction have been improved with our directed evolutionary phage display system. This novel phage display technology allows 12 copies of a specific desired peptide to be displayed on the exterior surface of Qβ in uniform distribution at the corners of the phage icosahedron. Through the recently optimized subtractive bio-panning strategy, fusion probes containing up to 80 amino acids altogether with linkers, can be displayed for target selection. Thus, combined uniqueness of its genome, structure, and proteins make the Qβ phage a desirable suitable innovation applicable in affinity maturation and directed evolutionary biotechnology. The evolutionary adaptability of the Qβ phage display strategy is still in its infancy. However, it has the potential to evolve functional domains of the desirable proteins, glycoproteins, and lipoproteins, rendering them superior to their natural counterparts.

scholarly journals Development of Two Murine Antibodies against Neospora caninum Using Phage Display Technology and Application on the Detection of N. caninum

PLoS ONE ◽  
2013 ◽  
Vol 8 (1) ◽  
pp. e53264 ◽  
Author(s):  
Jinhua Dong ◽  
Takahiro Otsuki ◽  
Tatsuya Kato ◽  
Tetsuya Kohsaka ◽  
Kazunori Ike ◽  
...  
Keyword(s):  
Phage Display ◽  
Neospora Caninum ◽  
Phage Display Technology ◽  
Display Technology

Production of Recombinant scFv against p24 of Human Immunodeficiency Virus Type 1 by Phage Display Technology

2014 ◽  
Vol 33 (1) ◽  
pp. 28-33 ◽  
Author(s):  
Sara Mohammadzadeh ◽  
Masoumeh Rajabibazl ◽  
Mehdi Fourozandeh ◽  
Mohammad Javad Rasaee ◽  
Fatemeh Rahbarizadeh ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Phage Display ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Phage Display Technology ◽  
Recombinant Scfv ◽  
Immunodeficiency Virus ◽  
Display Technology

Screening of peptide specific to cholangiocarcinoma cancer cells using an integrated microfluidic system and phage display technology

2017 ◽  
Vol 21 (9) ◽  
Author(s):  
Ching-Wen Yu ◽  
Chien-Yu Fu ◽  
Lien-Yu Hung ◽  
Chih-Hung Wang ◽  
Nai-Jung Chiang ◽  
...  
Keyword(s):  
Phage Display ◽  
Cancer Cells ◽  
Microfluidic System ◽  
Phage Display Technology ◽  
Display Technology
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