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

Abstract

Phage display technology serves as a powerful in vitro 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. Collec-tively, 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.