The genes coding for the variable (V) regions of the well-known murine anti-hen egg-white lysozyme (HEL) antibody D1.3 were inserted into pIII and displayed on the phage surface as fusion proteins. Subsequently, three groups showed that libraries of millions of random short peptides yielded specific peptides that could tightly bind to diverse antibodies used as selectors, thus laying the foundations for identifying specific peptides for any given antibody with no prior knowledge of its specificity.īy the beginning of the 1990s, John McCafferty and Gregory Winter extended the applications of the phage display methodology to antibody engineering. Three years later, Parmley and Smith described how peptide-phage fusions in a proportion as low as one peptide-phage fusion in a million wildtype virions could be enriched by using biotinylated antibodies specific for the peptides, a method coined as biopanning. Smith showed that foreign DNA fragments could be inserted into the gene encoding the filamentous phage coat protein III (pIII), expressed as peptide-phage fusions and enriched more than 1000-fold over wildtype phage by affinity to a specific antibody directed against the peptide. The first breakthrough in the remarkable development of phage display methodology, which became one of the most successful technology platforms for engineering antibody-based drugs, was published in 1985 by George Smith. The phage display methodology has revolutionized the protein engineering field, with a major impact on antibody engineering. Finally, we highlight current opportunities and challenges pertaining to phage display platforms and related display technologies. Second, we review the performance of the libraries in terms of the number of positive clones per panning, hit rate, affinity, and developability of the selected antibodies. We first discuss the quality of the libraries and the diverse types of antibody repertoires used as substrates to build the libraries, i.e., naïve, synthetic, and semisynthetic. Here, we compare nine phage display antibody libraries published in the last decade, which represent the state of the art in the discovery and development of therapeutic antibodies using phage display. This technology was initially developed by George Smith in the mid-1980s and applied by John McCafferty and Gregory Winter to antibody engineering at the beginning of 1990s. Phage display technology has played a key role in the remarkable progress of discovering and optimizing antibodies for diverse applications, particularly antibody-based drugs.
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