Phage Display as a Strategy to Obtain Anti-flavivirus Monoclonal Antibodies

Abstract

Arbovirus of the Flaviviridae family represents an issue worldwide, particularly because it can lead to serious illness and death in some countries. There is still a great complexity in obtaining effective therapies and specific and sensitive diagnostic tests, due to the high antigenic similarity between them. This similarity may account for antibodies cross reactivity which has positive and negative consequences for the course of infectious diseases. Among dengue virus (DENV) serotype infections, the cross-reactivity can increase virus replication and the risk of a severe disease by a mechanism known as an antibody-dependent enhancement (ADE). The search for serological biomarkers through monoclonal antibodies (MAbs) that identify unique viral regions can assist in the differential detection, whereas the development of recombinant antibodies with a neutralizing potential can lead to the establishment of efficacious treatments. The Phage Display methodology emerged as one of the main alternatives for the selection of human MAbs with high affinity for a specific target. Therefore, this technology can be a faster alternative for the development of specific diagnostic platforms and efficient and safe treatments for flavivirus infections. In this context, we propose for this chapter a discussion about Phage Display as a strategy to obtain MAbs for DENV and other flaviviruses.

Nanobodies: a promising approach to treatment of viral diseases

“…While biopanning and ELISA are valuable strategies for validating phage display libraries ( 25 ), there are some caveats and challenges associated with these techniques. Caveats for biopanning mostly include nonspecific binding, epitope masking, and limited diversity, while for ELISA, they encompass confirmation bias, false positives, and sensitivity issues.…”

“…One of the most common techniques for selecting proteins and antibodies is phage display ( 23 ). This technique has been widely employed due to its versatility, efficiency, and cost-effectiveness ( 16 , 17 , 24 , 25 ). To perform this selection, the protein of interest is presented on the surface of the phage, which then interacts with a diverse range of target molecules in a process known as biopanning ( 25 ).…”

“…This technique has been widely employed due to its versatility, efficiency, and cost-effectiveness ( 16 , 17 , 24 , 25 ). To perform this selection, the protein of interest is presented on the surface of the phage, which then interacts with a diverse range of target molecules in a process known as biopanning ( 25 ). This allows for the isolation and enrichment of specific ligands, particularly antigen-specific ligands in the case of antibodies ( 26 ).…”

Identification of Zika Virus NS1-Derived Peptides with Potential Applications in Serological Tests

“…Antibody detection assays targeting NS1 have been extensively utilized for ZIKV infection, although expressive cross-reactivity still occurs [ 53 ]. Although all peptides played a role in the immune recognition of NS1 in humans, the peptides ZKvROX4 and ZKvROX6 showed greater specificity for ZIKV while peptide ZKvROX5 showed cross-reactivity against DENV human sera, according to dot-blot results.…”

“…Indeed, serological assays based on NS1 protein have been used to detect previous infections by different flavivirus infections [ 20 ]. However, despite significant progress made in understanding ZIKV detection methods, further improvements are still needed [ 23 , 24 ].…”