Sanders, E.C.; Santos, A.M.; Nguyen, E.K.; Gelston, A.A.; Majumdar, S.; Weiss, G.A. Phage vs. Phage: Direct Selections of Sandwich Binding Pairs. Viruses2023, 15, 807.
Sanders, E.C.; Santos, A.M.; Nguyen, E.K.; Gelston, A.A.; Majumdar, S.; Weiss, G.A. Phage vs. Phage: Direct Selections of Sandwich Binding Pairs. Viruses 2023, 15, 807.
Sanders, E.C.; Santos, A.M.; Nguyen, E.K.; Gelston, A.A.; Majumdar, S.; Weiss, G.A. Phage vs. Phage: Direct Selections of Sandwich Binding Pairs. Viruses2023, 15, 807.
Sanders, E.C.; Santos, A.M.; Nguyen, E.K.; Gelston, A.A.; Majumdar, S.; Weiss, G.A. Phage vs. Phage: Direct Selections of Sandwich Binding Pairs. Viruses 2023, 15, 807.
Abstract
The sandwich format immunoassay is generally more sensitive and specific than more common assay formats, including direct, indirect, or competitive. A sandwich assay, however, requires two receptors to bind non-competitively to the target analyte. Typically, pairs of antibodies (Abs) or antibody fragments (Fabs) capable of forming a sandwiching with the target are identified through a slow, guess-and-check method with panels of candidate binding partners. Additionally, sandwich assays reliant on commercial antibodies can suffer from changes to reagent quality outside the researchers’ control. This report presents a reimagined and simplified phage display selection protocol that directly identifies sandwich binding peptides and Fabs. The approach yielded two sandwich pairs, one peptide-peptide and one Fab-peptide sandwich for the cancer and Parkinson’s disease biomarker DJ-1. Requiring just a few weeks to identify, the sandwich pairs delivered apparent affinity comparable to other commercial peptide and antibody sandwiches. The results reported here could expand the availability of sandwich binding partners for a wide range of clinical biomarker assays.
Chemistry and Materials Science, Analytical Chemistry
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
