Studying Binding Specificities of Peptide Recognition Modules by High-Throughput Phage Display Selections

2011 ◽  
pp. 87-97 ◽  
Author(s):  
Haiming Huang ◽  
Sachdev S. Sidhu
Keyword(s):  
Phage Display ◽  
High Throughput ◽  
Peptide Recognition

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 Creation of Novel Protein Transduction Domain (PTD) Mutants by a Phage Display-Based High-Throughput Screening System

2006 ◽  
Vol 29 (8) ◽  
pp. 1570-1574 ◽  
Author(s):  
Yohei Mukai ◽  
Toshiki Sugita ◽  
Tomoko Yamato ◽  
Natsue Yamanada ◽  
Hiroko Shibata ◽  
...  
Keyword(s):  
Phage Display ◽  
High Throughput ◽  
High Throughput Screening ◽  
Protein Transduction ◽  
Protein Transduction Domain ◽  
Screening System ◽  
Novel Protein

High-throughput screening of T7 phage display and protein microarrays as a methodological approach for the identification of IgE-reactive components

2018 ◽  
Vol 456 ◽  
pp. 44-53 ◽  
Author(s):  
Pablo San Segundo-Acosta ◽  
María Garranzo-Asensio ◽  
Carmen Oeo-Santos ◽  
Ana Montero-Calle ◽  
Joaquín Quiralte ◽  
...  
Keyword(s):  
Phage Display ◽  
High Throughput ◽  
High Throughput Screening ◽  
Methodological Approach ◽  
Protein Microarrays ◽  
T7 Phage Display ◽  
T7 Phage

Screening and Identification of PLK1-Polo Box Binding Peptides by High-Throughput Sequencing of Phage-Selected Libraries

2019 ◽  
Vol 26 (8) ◽  
pp. 620-633 ◽  
Author(s):  
Nousheen Bibi ◽  
Hafsa Niaz ◽  
Ted Hupp ◽  
Mohammad Amjad Kamal ◽  
Sajid Rashid
Keyword(s):  
Next Generation Sequencing ◽  
Phage Display ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
B Lymphocyte ◽  
Fluorescent Protein ◽  
Next Generation ◽  
Peptide Phage Display ◽  
Binding Peptides ◽  
Generation Sequencing

Background: Human proteome contains a plethora of short linear peptide motifs that is crucial for signaling and other cellular processes. These motifs are difficult to identify due to lack of systematic approach for their detection. Objective: Here we demonstrate the use of peptide phage display in combination with high throughput next generation sequencing to identify enriched peptide sequences through biopanning process against polo box domain (PBD) of mitotic polo like kinase 1 (Plk1). Methods: Purified recombinant Plk1 and two unrelated controls namely B-lymphocyte antigen (CD20) and fluorescent protein (mCherry) were subjected to peptide phage display analysis. Bacterially-propagated phage DNA was amplified by PCR using triplet bar coded primers to tag the pool from each amplicon. Results: Proteomic peptide phage display along with next generation sequencing and Bioinformatics analysis demonstrated several known and putative novel interactions which were potentially related to Plk1-PBD. With our strategy, we were able to identify and characterize several Plk1-PBD binding peptides, as well as define more precisely, consensus sequences. Conclusion: We believe that this information could provide valuable tools for exploring novel interaction involved in Plk1 signaling as well as to choose peptides for Plk1 specific drug development.

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

scholarly journals High-throughput Pyrosequencing™ of a phage display library for the identification of enriched target-specific peptides

BioTechniques ◽  
2003 ◽  
Vol 35 (2) ◽  
pp. 317-324 ◽  
Author(s):  
Ahad Rahim ◽  
Charles Coutelle ◽  
Richard Harbottle
Keyword(s):  
Phage Display ◽  
High Throughput ◽  
Phage Display Library

scholarly journals A novel approach for characterisation of conformational allergen epitopes combining phage display and high-throughput sequencing

2014 ◽  
Vol 4 (S2) ◽  
Author(s):  
Anders Christiansen ◽  
Christian Skjodt Hansen ◽  
Jens Vindahl Kringelum ◽  
Ole Lund ◽  
Katrine Lindholm Bogh ◽  
...  
Keyword(s):  
Phage Display ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Novel Approach
Sign in / Sign up

Export Citation Format

Share Document