title: Real-time analysis of dual-display phage immobilization and autoantibody screening using quartz crystal microbalance with dissipation monitoring.
abstract: Over the last three decades, phage display technology has been used for the display of target-specific biomarkers, peptides, antibodies, etc. Phage display-based assays are mostly limited to the phage ELISA, which is notorious for its high background signal and laborious methodology. These problems have been recently overcome by designing a dual-display phage with two different end functionalities, namely, streptavidin (STV)-binding protein at one end and a rheumatoid arthritis-specific autoantigenic target at the other end. Using this dual-display phage, a much higher sensitivity in screening specificities of autoantibodies in complex serum sample has been detected compared to single-display phage system on phage ELISA. Herein, we aimed to develop a novel, rapid, and sensitive dual-display phage to detect autoantibodies presence in serum samples using quartz crystal microbalance with dissipation monitoring as a sensing platform. The vertical functionalization of the phage over the STV-modified surfaces resulted in clear frequency and dissipation shifts revealing a well-defined viscoelastic signature. Screening for autoantibodies using antihuman IgG-modified surfaces and the dual-display phage with STV magnetic bead complexes allowed to isolate the target entities from complex mixtures and to achieve a large response as compared to negative control samples. This novel dual-display strategy can be a potential alternative to the time consuming phage ELISA protocols for the qualitative analysis of serum autoantibodies and can be taken as a departure point to ultimately achieve a point of care diagnostic system.
authors: Biomedical Research Institute (BIOMED), Hasselt University, Hasselt, Belgium.
authors: Kaushik
authors: K
authors: Rajaram
authors: Institute for Materials Research (IMO), Hasselt University, Diepenbeek, Belgium ; IMEC vzw, Division IMOMEC, Diepenbeek, Belgium.
authors: Patricia
authors: P
authors: Losada-Pérez
authors: Biomedical Research Institute (BIOMED), Hasselt University, Hasselt, Belgium.
authors: Veronique
authors: V
authors: Vermeeren
authors: Biomedical Research Institute (BIOMED), Hasselt University, Hasselt, Belgium.
authors: Baharak
authors: B
authors: Hosseinkhani
authors: Institute for Materials Research (IMO), Hasselt University, Diepenbeek, Belgium ; Soft Matter and Biophysics Section, Department of Physics and Astronomy, KU Leuven, Leuven, Belgium.
authors: Patrick
authors: P
authors: Wagner
authors: Biomedical Research Institute (BIOMED), Hasselt University, Hasselt, Belgium.
authors: Veerle
authors: V
authors: Somers
authors: Biomedical Research Institute (BIOMED), Hasselt University, Hasselt, Belgium.
authors: Luc
authors: L
authors: Michiels
mesh_terms: D001172:Arthritis, Rheumatoid / Q000276:immunology; D001323:Autoantibodies / Q000737:chemistry*; D001435:Bacteriophages; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D019151:Peptide Library; D010455:Peptides; D011485:Protein Binding; D058525:Quartz Crystal Microbalance Techniques; D019809:Streptavidin / Q000737:chemistry; D020349:Surface Plasmon Resonance
chemical_list: D001323:Autoantibodies; D007074:Immunoglobulin G; D019151:Peptide Library; D010455:Peptides; D019809:Streptavidin
keywords: dissipation monitoring; dual-display phage; quartz crystal microbalance; rheumatoid arthritis; streptavidin-binding protein; surface plasmon resonance