Construction and Applications of Mammalian Cell-Based DNA-Encoded Peptide/Protein Libraries

Abstract

DNA-encoded peptide/protein libraries are the starting point for protein evolutionary modification and functional peptide/antibody selection. Different display technologies, protein directed evolution, and deep mutational scanning (DMS) experiments employ DNA-encoded libraries to provide sequence variations for downstream affinity-or function-based selections. Mammalian cells promise the inherent post-translational modification and near-to-natural conformation of exogenously expressed mammalian proteins and thus are the best platform for studying transmembrane proteins or human disease-related proteins. However, due to the current technical bottlenecks of constructing mammalian cell-based large size DNA-encoded libraries, the advantages of mammalian cells as screening platforms have not been fully exploited. In this review, we summarize the current efforts in constructing DNA-encoded libraries in mammalian cells and the existing applications of these libraries in different fields.

Preclinical Anticipation of On- and Off-Target Resistance Mechanisms to Anti-Cancer Drugs: A Systematic Review

“…One strategy to enable the systematic discovery of on-target resistance mutations is DMS, which, unlike the methods described above, is a very comprehensive approach based on specifically designed libraries (Table 1). All possible single amino acid substitutions at all positions of the protein of interest are covered and the corresponding lentivirus expression library is then transduced into appropriate cells [14,45,73,74]. Functional assays to determine target protein binding to other biomolecules, stability, or activity can then be performed.…”

“…More recently, systematic strategies have been developed for an unbiased approach. One example is detailed mutational scanning of the drug target to determine which positions are essential for oncogenic activity or associated with treatment resistance [14]. A major limitation of this procedure is that it only addresses on-target and not off-target resistance mechanisms.…”

What is the role of microbial biotechnology and genetic engineering in medicine?

“…On the other hand, if a hybridoma cell line is used, the antibody can be made recombinant by sequencing the DNA of the hybridoma cell line and subsequently cloning a gene encoding the identified sequence (Andrews et al, 2019 ). Finally, another method to produce recombinant antibodies is by selecting antigen binding from recombinant antibody libraries (Wang et al, 2023 ). Chimeric and humanized antibodies are two types of nonhuman recombinant antibodies whose sequences have been obtained from the nonhuman immune system (such as from mice).…”