scholarly journals Practical Tips for Construction of Custom Peptide Libraries and Affinity Selection by Using Commercially Available Phage Display Cloning Systems

2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Keisuke Fukunaga ◽  
Masumi Taki
Keyword(s):  
Phage Display ◽  
Basic Amino Acid ◽  
Phage Library ◽  
Affinity Selection ◽  
Phage Display Technology ◽  
Display Technology ◽  
Phage Libraries ◽  
Type 3 ◽  
T7 Phage ◽  
Selection Of

Phage display technology is undoubtedly a powerful tool for affinity selection of target-specific peptide. Commercially available premade phage libraries allow us to take screening in the easiest way. On the other hand, construction of a custom phage library seems to be inaccessible, because several practical tips are absent in instructions. This paper focuses on what should be born in mind for beginners using commercially available cloning kits (Ph.D. with type 3 vector and T7Select systems for M13 and T7 phage, respectively). In the M13 system, Pro or a basic amino acid (especially, Arg) should be avoided at the N-terminus of peptide fused to gp3. In both systems, peptides containing odd number(s) of Cys should be designed with caution. Also, DNA sequencing of a constructed library before biopanning is highly recommended for finding unexpected bias.

Efficient one-cycle affinity selection of binding proteins or peptides specific for a small-molecule using a T7 phage display pool

2008 ◽  
Vol 16 (22) ◽  
pp. 9837-9846 ◽  
Author(s):  
Yoichi Takakusagi ◽  
Kouji Kuramochi ◽  
Manami Takagi ◽  
Tomoe Kusayanagi ◽  
Daisuke Manita ◽  
...  
Keyword(s):  
Phage Display ◽  
Small Molecule ◽  
Binding Proteins ◽  
Affinity Selection ◽  
T7 Phage Display ◽  
T7 Phage ◽  
Selection Of

A novel phage display vector for selection of target-specific peptides

2020 ◽  
Vol 33 ◽  
Author(s):  
Alex Chang ◽  
Joey P Ting ◽  
Alfonso Espada ◽  
Howard Broughton ◽  
Manuel Molina-Martin ◽  
...  
Keyword(s):  
Amino Acids ◽  
Fusion Protein ◽  
Phage Display ◽  
Copy Number ◽  
Wild Type ◽  
Phage Display Technology ◽  
Successful Isolation ◽  
Display Technology ◽  
Therapeutic Peptides ◽  
Selection Of

Abstract Intrinsic low display level of polypeptides on phage is a fundamental and limiting hurdle in successful isolation of target-specific binders by phage display technology. To circumvent this challenge, we optimized the copy number of peptides displayed on the phage surface using type 33 phage vector. We randomized the first 67 amino acids of the wild type PIII to identify mutants that would result in its reduced expression. Consequently, the display level was improved by 30-fold due to higher incorporation of the synthetic PIII–peptide fusion protein on the phage surface. Utilization of this novel phage vector should provide a solid basis for the discovery of therapeutic peptides.

scholarly journals Mimotopes for Mycotoxins Diagnosis Based on Random Peptides or Recombinant Antibodies from Phage Library

Molecules ◽  
2021 ◽  
Vol 26 (24) ◽  
pp. 7652
Author(s):  
Wei Sun ◽  
Yan Zhang ◽  
Zhigang Ju
Keyword(s):  
Phage Display ◽  
Recombinant Antibody ◽  
Recombinant Antibodies ◽  
Phage Display Library ◽  
Ease Of Use ◽  
Phage Library ◽  
Phage Display Technology ◽  
Random Peptide ◽  
Display Technology

Mycotoxins, the small size secondary metabolites of fungi, have posed a threat to the safety of medicine, food and public health. Therefore, it is essential to create sensitive and effective determination of mycotoxins. Based on the special affinity between antibody and antigen, immunoassay has been proved to be a powerful technology for the detection of small analytes. However, the tedious preparation and instability of conventional antibodies restrict its application on easy and fast mycotoxins detection. By virtue of simplicity, ease of use, and lower cost, phage display library provides novel choices for antibodies or hapten conjugates, and lead random peptide or recombinant antibody to becoming the promising and environmental friendly immune-reagents in the next generation of immunoassays. This review briefly describes the latest developments on mycotoxins detection using M13 phage display, mainly focusing on the recent applications of phage display technology employed in mycotoxins detection, including the introduction of phage and phage display, the types of phage displayed peptide/recombinant antibody library, random peptides/recombinant antibodies-based immunoassays, as well as simultaneous determination of multiple mycotoxins.

scholarly journals Selection of Linkers for a Catalytic Single-chain Antibody Using Phage Display Technology

1996 ◽  
Vol 271 (26) ◽  
pp. 15682-15686 ◽  
Author(s):  
Ying Tang ◽  
Ning Jiang ◽  
Cushrow Parakh ◽  
Donald Hilvert
Keyword(s):  
Phage Display ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Phage Display Technology ◽  
Display Technology ◽  
Selection Of

K-Ras(G12D)-selective inhibitory peptides generated by random peptide T7 phage display technology

2017 ◽  
Vol 484 (3) ◽  
pp. 605-611 ◽  
Author(s):  
Kotaro Sakamoto ◽  
Yusuke Kamada ◽  
Tomoya Sameshima ◽  
Masahiro Yaguchi ◽  
Ayumu Niida ◽  
...  
Keyword(s):  
Phage Display ◽  
Phage Display Technology ◽  
Random Peptide ◽  
Inhibitory Peptides ◽  
Display Technology ◽  
T7 Phage Display ◽  
T7 Phage

scholarly journals Use of phage display technology for the selection of monoclonal antibodies in allergen research and diagnostics

2014 ◽  
Vol 4 (S2) ◽  
Author(s):  
Marjolein Vandekerckhove ◽  
Bart Van Droogenbroeck ◽  
Marc Deloose ◽  
Hilde Lapeere ◽  
Gevaert Philippe
Keyword(s):  
Monoclonal Antibodies ◽  
Phage Display ◽  
Phage Display Technology ◽  
Display Technology ◽  
Selection Of

scholarly journals Phage display as a tool for identifying HIV-1 broadly neutralizing antibodies

2021 ◽  
Vol 25 (5) ◽  
pp. 562-572
Author(s):  
A. N. Chikaev ◽  
A. P. Rudometov ◽  
Yu. A. Merkulyeva ◽  
L. I. Karpenko
Keyword(s):  
Phage Display ◽  
High Throughput Screening ◽  
Neutralizing Antibodies ◽  
Selection Procedure ◽  
Broadly Neutralizing Antibodies ◽  
Recombinant Phage ◽  
Phage Display Technology ◽  
Display Technology ◽  
Phage Libraries ◽  
Hiv 1

Combinatorial biology methods offer a good solution for targeting interactions of specific molecules by a high-throughput screening and are widely used for drug development, diagnostics, identification of novel monoclonal antibodies, search for linear peptide mimetics of discontinuous epitopes for the development of immunogens or vaccine components. Among all currently available techniques, phage display remains one of the most popular approaches. Despite being a fairly old method, phage display is still widely used for studying protein-protein, peptide-protein and DNA-protein interactions due to its relative simplicity and versatility. Phage display allows highly representative libraries of peptides, proteins or their fragments to be created. Each phage particle in a library displays peptides or proteins fused to its coat protein and simultaneously carries the DNA sequence encoding the displayed peptide/protein in its genome. The biopanning procedure allows isolation of specific clones for almost any target, and due to the physical link between the genotype and the phenotype of recombinant phage particles it is possible to determine the structure of selected molecules. Phage display technology continues to play an important role in HIV research. A major obstacle to the development of an effective HIV vaccine is an extensive genetic and antigenic variability of the virus. According to recent data, in order to provide protection against HIV infection, the so-called broadly neutralizing antibodies that are cross-reactive against multiple viral strains of HIV must be induced, which makes the identification of such antibodies a key area of HIV vaccinology. In this review, we discuss the use of phage display as a tool for identification of HIV-specific antibodies with broad neutralizing activity. We provide an outline of phage display technology, briefly describe the design of antibody phage libraries and the affinity selection procedure, and discuss the biology of HIV-1-specific broadly neutralizing antibodies. Finally, we summarize the studies aimed at identification of broadly neutralizing antibodies using various types of phage libraries.

scholarly journals Improved affinity selection using phage display technology and off-rate based selection

2006 ◽  
Vol 9 (2) ◽  
pp. 171-175 ◽  
Author(s):  
Bin Yuan ◽  
Philip Schulz ◽  
Ruitian Liu ◽  
Michael R. Sierks
Keyword(s):  
Phage Display ◽  
Affinity Selection ◽  
Phage Display Technology ◽  
Display Technology
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