Selection of phage-displayed antibodies with high affinity and specificity by electrophoresis in microfluidic devices

SANLUIS-VERDES, ANAHÍ; PEÑAHERRERA, ANA; TORÁN, JOSÉ L.; ROSERO, GUSTAVO; NORIEGA, MARÍA A.; LERNER, BETIANA; CASASNOVAS, JOSÉ M.; PÉREZ, MAXIMILIANO

ELECTROPHORESIS

WILEY-V C H VERLAG GMBH

Año: 2023

0173-0835

A method development aimed for high-throughput and automated antibody screening holds great potential for areas ranging from fundamental molecular interactions to the discovery of novel disease markers, therapeutic targets, and monoclonal antibody engineering. Surface display techniques enable efficient manipulation of large molecular libraries in small volumes. Specifically, phage display appeared as a powerful technology for selecting peptides and proteins with enhanced, target-specific binding affinities. Here, we present a phage-selection microfluidic device wherein electrophoresis was performed under two orthogonal electric fields through an agarose gel functionalized with the respective antigen. This microdevice was capable of screening and sorting in a single round high-affinity phage-displayed antibodies against virus glycoproteins, including human immunodeficiency virus-1 glycoprotein 120 or the Ebola virus glycoprotein (EBOV-GP). Phages were differentially and laterally swept depending on their antigen affinity; the high-affinity phages were recovered at channels proximal to the application site, whereas low-affinity phages migrated distal after electrophoresis. These experiments proved that the microfluidic device specifically designed for phage-selection is rapid, sensitive, and effective. Therefore, this is an efficient and cost-effective method that allowed highly controlled assay conditions for isolating and sorting high-affinity ligands displayed in phages.