Phage Display Technology as a Powerful Platform for Antibody Drug Discovery

Phage Display Technology

Phage display technology, allowing the construction of libraries in which various peptides and proteins are displayed on the phage, has widespread applications.

Localization of antigenic determinant clusters
Identification of protein interaction sites
Isolation of specific regulatory molecules for the preparation of artificial antibodies and vaccines
Diagnostic techniques
Research and development of enzyme inhibitors
Peptide drug development
Antibody drug development

Fig 1. Schematic representation of different types of antibody formats displayed on the phages. (Kazuya, et al. 2021)

Antibody Drug Development Based on Phage Display Technology

The use of hybridomas to manufacture monoclonal antibodies was invented in 1975. Because of their great specificity and affinity for the target molecule, monoclonal antibodies have been considered a possible "panacea," but clinical studies have proven disappointing. There are several causes of failure, many of which can be addressed.

Development of Human Monoclonal Antibodies

Use phage display-derived human antibody library to screen for suitable antibodies.
Humanization of mouse monoclonal antibodies using the guided selection approach based on phage display technology.

Improving the Affinity of Monoclonal Antibodies

Gene libraries, constructed using mutations in antigen-binding regions such as VL and VH, are displayed on phage, and antibodies with high affinity for the antigen are then isolated by biopanning.

Methods for constructing gene libraries with mutations:

Random mutagenesis
Site-specific mutagenesis

Target Discovery Based on Phage Display Technology

Despite their distinct and superior properties, antibody drugs' targets are dwindling. As a result, identifying prospective targets for antibody drugs is critical. The use of phages to identify possible therapeutic targets is an efficient method.

Phage Display-Based Technology to Screen for Antigens That React With Autoantibodies

Autoantibodies to autoantigens can be employed as biomarkers in autoimmune disorders. The search for antigens that bind to autoantibodies in autoimmune diseases is a potential strategy for elucidating molecular pathophysiology and target discovery, and phage display techniques can be useful in studying autoimmune-specific autoantigens.

Construct a complementary DNA (cDNA) phage display library based on the mRNA extracted from biological tissues
Phage clones bound to antibodies collected from the blood of patients with autoimmune diseases and other sources are enriched by a biopanning of this library
Identify autoantigens by analyzing cDNA sequences in eluted phages using high-throughput techniques

High-Throughput Validation of Therapeutic Target Candidates Based on Phage Display Technology

Antibody proteomics technology isolates monoclonal antibodies to candidate proteins using phage antibody libraries and TMA analysis to find potential biomarkers and therapeutic targets.

Search for disease-related proteins by proteomics-based analysis using 2D-DIGE
Identification of candidate proteins using mass spectrometry
Isolation of monoclonal antibodies against candidate proteins using phage antibody libraries
Validation of candidate proteins using tissue microarray staining analysis

Fig 2. Schematic illustration of the antibody proteomics system. (Kazuya, et al. 2021)

Reference

Nagano, K.Tsutsumi, Y., Phage display technology as a powerful platform for antibody drug discovery. Viruses, 2021. 13(2).

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