Library-based display technologies: where do we stand?

Abstract

Over the past two decades, library-based display technologies have been staggeringly optimized since their appearance in order to mimic the process of natural molecular evolution. Display technologies are essential for the isolation of specific high-affinity binding molecules (proteins, polypeptides, nucleic acids and others) for diagnostic and therapeutic applications in cancer, infectious diseases, autoimmune, neurodegenerative, inflammatory pathologies etc. Applications extend to other fields such as antibody and enzyme engineering, cell-free protein synthesis and the discovery of protein-protein interactions. Phage display technology is the most established of these methods but more recent fully in vitro alternatives, such as ribosome display, mRNA display, cis-activity based (CIS) display and covalent antibody display (CAD), as well as aptamer display and in vitro compartmentalization, offer advantages over phage in library size, speed and the display of unnatural amino acids and nucleotides. Altogether, they have produced several molecules currently approved or in diverse stages of clinical or preclinical testing and have provided researchers with tools to address some of the disadvantages of peptides and nucleotides such as their low affinity, low stability, high immunogenicity and difficulty to cross membranes. In this review we assess the fundamental technological features and point out some recent advances and applications of display technologies.

Peptidomimetic therapeutics: scientific approaches and opportunities

“…Unnatural amino acid-containing peptidomimetics can be developed by single-residue scanning of a parent sequence or scaffold [14], computational and rational design, or phenotype and/or genotype-linked display from phage [23] or yeast [24]. N -Alkyl amino acids (e.g.…”

“…Using advances in molecular evolution and display, investigators rapidly selected high-affinity candidates from their library of 147 noncanonical and unnatural amino acids. Phage display is well established, and there are also other techniques, such as yeast display, based on combinatorial molecular display or in vitro compartmentalization, which are amenable to the display of unnatural amino acids [23]. …”

Sustainable therapies by engineered bacteria

“…In this context, microbial biotechnology already provides important solutions at various levels, such as increasing production yields in bioreactors using microorganisms (i.e. yeasts, bacteria), reducing the time and cost associated with the slow growth of mammalian cells, and assisting on the optimization and engineering of biologics thanks to the high cloning capacity of microorganisms and the efficient screening methods of microbial libraries (Hoogenboom, 2005; Galan et al ., 2016). In addition to these conventional technologies, microbial engineering is developing innovative approaches to produce and deliver complex biologics in situ , within the body, ideally in a controlled and programmable way.…”

High Affinity Maturated Human Antibodies from Naïve and Synthetic Antibody Repertoires

“…Puromycin functions to mimic the role of amino-acyl tRNA by attaching itself to a DNA primer affixed to the mRNA template. This allows puromycin to attach itself covalently to the nascent antibody protein based on the peptidyl transferase activity of the ribosome [12,22].…”

“…The covalent display technology (CDT) exploits the properties of the replication initiator protein from E. coli bacteriophage P2 [31]. A pool of DNA encoding antibody molecules is generated as a fusion to the P2A coding sequence [12]. The DNA pool is then transcribed and translated using cell free expression systems.…”