Programmable Nucleic Acid Nanoswitches for the Rapid, Single-Step Detection of Antibodies in Bodily Fluids

Abstract

Antibody detection plays a pivotal role in the diagnosis of pathogens and monitoring the success of vaccine immunization. However, current serology techniques require multiple, time-consuming washing and incubation steps, which limit their applicability in point-of-care (POC) diagnostics and high-throughput assays. We developed here a nucleic acid nanoswitch platform able to instantaneously measure immunoglobulins of type G and E (IgG and IgE) levels directly in blood serum and other bodily fluids. The system couples the advantages of target-binding induced colocalization and nucleic acid conformational-change nanoswitches. Due to the modular nature of the recognition platform, the method can potentially be applied to the detection of any antibody for which an antigen can be conjugated to a nucleic acid strand. In this work we show the sensitive, fast and cost-effective detection of four different antibodies and demonstrate the possible use of this approach for the monitoring of antibody levels in HIV+ patients immunized with AT20 therapeutic vaccine.

A Universal Proximity CRISPR Cas12a Assay for Ultrasensitive Detection of Nucleic Acids and Proteins

“…As sensitive detection of antibodies in clinical samples plays critical roles for the diagnosis of infectious and autoimmune diseases, [7][8][9] we chose anti-biotin antibody as a testbed for our assay. Correspondingly, biotin was used as the affinity ligand ( Fig.…”

Recent Developments of Electrochemical and Optical Biosensors for Antibody Detection

“…Using the base-pairing chemistry of DNA, optical-based biosensors allow for accurate and rapid antibody detection. Recent research demonstrated a sensitive, and highly specific method to detect antibodies ( Figure 7a) [60]. The recognition element was the epitope or corresponding antigen to the antibody of interest.…”

“…The signal gain is recorded with different concentrations. (Adapted with permission from Ref [60]. Copyright 2018 American Chemical Society.)…”

Decorating bacteria with self-assembled synthetic receptors

“…In recent years, considerable attention has been devoted to developing protein binders based on oligodeoxynucleotide (ODN)-small-molecule conjugates [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27] that, similar to CSPs, can respond to external stimuli and undergo dynamic structural changes that enable them to reversibly interact with proteins and regulate their functions [11][12][13][14][15][16][17][18][19][20][21] . We have recently developed protein binders based on ODN-small-molecule conjugates [21][22][23] and have shown that such binders can be designed to reversibly interact with different proteins 21,22 and even mediate unnatural protein-protein communication processes 21 .…”

“…Inspired by recent successes in dynamically controlling protein functions and binding interactions with ODN-small-molecule conjugates [11][12][13][14][15][16][17][18][19][20][21] and in cell surface engineering , we set out to develop a dynamic artificial receptor system that combines principles from both research areas. Specifically, we hypothesized that by attaching DNA-based, stimuli-responsive protein binders on the surfaces of living bacteria, at a specific position, and by using solely self-assembly processes, a biomimetic system that imitates the dynamic features of natural CSPs can be obtained.…”