Considerations on Antibody-Phage Display Methodology

Abstract

For almost 15 years phage display is being used for the selection of specific antigen-binders from artificial libraries of single chain antibodies. Filamentous phages have been developed in a way to express foreign proteins on the surface and at the same time carrying the genetic information of the surface expressed molecule within the phage capsid. This property guarantees the coupling of phenotype and genotype during phage amplification and affinity selection. The possibility to generate large antibody libraries and the simplified antibody-backbone of a single chain antibody has made antibody-phage display to a powerful tool for the development of new therapeutics against various human diseases. In this review we discuss the general principles and latest developments and applications in antibody phage display technology.

Production of a phage-displayed single chain variable fragment antibody against infectious bursal disease virus

“…Phage display technology provides a straightforward approach for selection of specific recombinant antibody fragments with highest affinity for the target antigen from a huge number of variants in antibody libraries [19]. On that basis, in the present study, this technology was applied to isolate new antibodies against the IBDV virions from Tomlinson I naïve nonimmunized antibody libraries.…”

Progress in phage display: evolution of the technique and its applications

“…Applications of phage display have been extensively reviewed elsewhere [23,[114][115][116][117][118][119][120][121][122][123][124][125][126][127][128][129][130][131][132] and will not be discussed here in detail. The technology, often requiring creative modifications, was used for epitope mapping of antibodies [114], screening for receptor agonists and antagonists [115,133,134], in vitro evolution of antibodies [116,117] and antibody surrogates in the form of randomized fragments on diverse scaffold proteins [118,119], discovery of enzyme substrates [120,121] and inhibitors [113,122,135], identifying functionally coupled proteins and analysis of protein-protein interactions [123,124], designing catalytic antibodies (abzymes) and enzymes with novel specificities [125], improving proteolytic and folding stability of muteins [126], vaccine design [131,132], and construction of gene delivery vehicles [127].…”

“…The technology, often requiring creative modifications, was used for epitope mapping of antibodies [114], screening for receptor agonists and antagonists [115,133,134], in vitro evolution of antibodies [116,117] and antibody surrogates in the form of randomized fragments on diverse scaffold proteins [118,119], discovery of enzyme substrates [120,121] and inhibitors [113,122,135], identifying functionally coupled proteins and analysis of protein-protein interactions [123,124], designing catalytic antibodies (abzymes) and enzymes with novel specificities [125], improving proteolytic and folding stability of muteins [126], vaccine design [131,132], and construction of gene delivery vehicles [127]. Protein ligands selected from phage libraries as well as whole recombinant phage have been used as affinity matrices in chromatographic techniques [128,129] and biosensing [130].…”

Selection of proteins with desired properties from natural proteome libraries using mRNA display

“…Phage display currently allows a complexity in the range of 10 9 -10 10 (ref. 12). The library size for cell-based selections, such as the yeast two-hybrid system, bacteria and yeast surface display and so on, is typically limited to ~1 million 19 .…”