Evaluation of Phage Display Biopanning Strategies for the Selection of Anti-Cell Surface Receptor Antibodies

Abstract

Monoclonal antibodies (mAbs) are one of the most successful and versatile protein-based pharmaceutical products used to treat multiple pathological conditions. The remarkable specificity of mAbs and their affinity for biological targets has led to the implementation of mAbs in the therapeutic regime of oncogenic, chronic inflammatory, cardiovascular, and infectious diseases. Thus, the discovery of novel mAbs with defined functional activities is of crucial importance to expand our ability to address current and future clinical challenges. In vitro, antigen-driven affinity selection employing phage display biopanning is a commonly used technique to isolate mAbs. The success of biopanning is dependent on the quality and the presentation format of the antigen, which is critical when isolating mAbs against membrane protein targets. Here, we provide a comprehensive investigation of two established panning strategies, surface-tethering of a recombinant extracellular domain and cell-based biopanning, to examine the impact of antigen presentation on selection outcomes with regards to the isolation of positive mAbs with functional potential against a proof-of-concept type I cell surface receptor. Based on the higher sequence diversity of the resulting antibody repertoire, presentation of a type I membrane protein in soluble form was more advantageous over presentation in cell-based format. Our results will contribute to inform and guide future antibody discovery campaigns against cell surface proteins.

Production and Characterization of Monoclonal antibodies to Xenopus proteins

“…Another approach that has been used extensively in the pharmaceutical industry is the use of recombinant antibodies (Maruthachalam et al, 2022; Panagides et al, 2022). Libraries of variable chain antibodies exist that can be used to screen and identify binders.…”

“…Another approach that has been used extensively in the pharmaceutical industry is the use of recombinant antibodies (Maruthachalam et al, 2022;Panagides et al, 2022).…”

Selection of phage display peptides against Pasteurella multocida using suspension method of biopanning

“…Generally 3 rounds of selection are sufficient but more number of selection rounds could be employed to increase the pool of more specific phages (Hyman 2019). In this study, the suspension method of selection was followed over the surface panning procedure to overcome the limitation of selecting non-specific phages to BSA in format (Panagides et al 2022). However, in the suspension format of selection, non-specific binding phages to the other proteins and surfaces could be avoided.…”

Computer-Assisted Design of Peptide-Based Radiotracers

“…Biopanning [ 22 ] was used as a screening method [ 23 , 24 ] to select the Aβ binding peptide probe (ABPP) sequences. After four rounds of biopanning performed in different conditions, each screened phage presenting its peptide sequence was analyzed, and the best one was selected.…”