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.

Harnessing phage display for the discovery of peptide-based drugs and monoclonal antibodies

2020 ◽  
Vol 27 ◽  
Author(s):  
Yuyang Li ◽  
Min Liu ◽  
Songbo Xie
Keyword(s):  
Monoclonal Antibodies ◽  
Drug Discovery ◽  
Phage Display ◽  
High Throughput Screening ◽  
Phage Display Technology ◽  
Future Directions ◽  
Drug Discovery And Development ◽  
Display Technology ◽  
Phage Capsid ◽  
Functional Peptides

: Phage display is a powerful high-throughput screening technology that presents a large and diverse selection of functional peptides, proteins, or antibody fragments on phage capsid surfaces for affinity determination towards a target of interest. Due to its advantages of a large screening capacity, mass production through fermentation, and straightforward execution, phage display has been widely used in bioengineering and biomedicine, especially for diagnostics and therapeutics. With the advent of next-generation sequencing and microfluidics technologies, phage display has become an even more powerful and popular tool for drug discovery and development. Here, we briefly review phage display technology and its application to drug discovery, including the discovery and development of peptide-based drugs and monoclonal antibodies. Combined with the emerging proteolysis targeting chimeras (PROTAC), we also discuss the advantages and future directions, aiming to facilitate the drug discovery and development.

scholarly journals Semi-Automated Cell Panning for Efficient Isolation of FGFR3-Targeting Antibody

2021 ◽  
Vol 22 (12) ◽  
pp. 6240
Author(s):  
Byeongkwi Min ◽  
Minyoung Yoo ◽  
Hyeree Kim ◽  
Minjung Cho ◽  
Do-Hyun Nam ◽  
...  
Keyword(s):  
Membrane Protein ◽  
High Throughput Screening ◽  
Magnetic Beads ◽  
Conformational Structure ◽  
Single Chain ◽  
Screening Process ◽  
Phage Display Technology ◽  
Display Technology ◽  
Cell Membrane Protein ◽  
Phage Libraries

Phage display technology is a widely used practical tool for isolating binding molecules against the desired targets in phage libraries. In the case of targeting the membrane protein with its natural conformation, conventional bio-panning has limitations on the efficient screening of the functionally relevant antibodies. To enrich the single-chain variable fragment (scFv) pools for recognizing the natural conformation of the membrane targets, the conventional bio-panning and screening process was modified to include the semi-automated cell panning protocol. Using FGFR3-overexpressing patient-derived cancer cells, biotin-X-DHPE was introduced and coupled to Streptavidin-coated magnetic beads for use in the solution-phage bio-panning procedure. The resulting clones of scFv were compared to the diversity of the binding region, especially on CDR-H3. The clones enriched further by cell-based panning procedure possessed a similar binding site and the CDR-H3 loop structure. The resulting antibodies inhibited cell growth and induced target degradation. This process may be a useful tool for screening biologically related antibodies that recognize natural conformational structure on cell membrane protein. Furthermore, cell-based panning has the potential to further expand to a high-throughput screening (HTS) system and automation process.

scholarly journals Identification of a LFA-1 region involved in the HIV-1-induced syncytia formation through phage-display technology

2001 ◽  
Vol 31 (1) ◽  
pp. 57-63 ◽  
Author(s):  
Francesca Poloni ◽  
Patrizia Puddu ◽  
Franca Moretti ◽  
Michela Flego ◽  
Giulia Romagnoli ◽  
...  
Keyword(s):  
Phage Display ◽  
Phage Display Technology ◽  
Display Technology ◽  
Syncytia Formation ◽  
Hiv 1

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.

Broadly Neutralizing Antibodies (BNAbs): templates for HIV-1 vaccine design

2017 ◽  
Vol 2 (4) ◽  
Author(s):  
Lixin Yan ◽  
◽  
Lihong Liu ◽  
Yilin Wang ◽  
Xi Huang ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Vaccine Design ◽  
Broadly Neutralizing Antibodies ◽  
Hiv 1

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 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.

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