Phage Display Technology: Design, Library Construction, and Panning Strategies for Antibody Development

Phage display technology serves as a powerful <i>in vitro</i> platform for discovering antigen-specific antibodies, enabling the rapid identification and engineering of monoclonal antibodies for therapeutic applications. This review discusses key factors such as phagemid vector, antibody fragment formats, in-frame selection strategies, and effective library size in the context of maximizing functional diversity. Based on these factors, a detailed comparison of phage display antibody libraries is provided, including naïve, synthetic, and semi-synthetic formats, with emphasis on their distinctive features and design strategies. Furthermore, major panning strategies-solid-phase, liquid-phase, and cell-based panning-are compared, highlighting their respective strengths and limitations for isolating functional binders with regard to antigen presentation, epitope accessibility, and selection fidelity. Collectively, the refinement of library design and selection methodologies has led to the development of numerous approved antibodies, establishing phage display as a foundational technology in antibody drug discovery. Ultimately, this review aims to facilitate the effective application of phage display technology in developing antibody-based therapeutics.