TY - JOUR
T1 - Yeast Surface-Displayed Quenchbody as a Novel Whole-Cell Biosensor for One-Step Detection of Influenza A (H1N1) Virus
AU - Seo, Yoonjoo
AU - Zhou, Aijia
AU - Nguyen, Thanh H.
AU - Wei, Na
N1 - This work was supported by the Strategic Research Initiatives Program at the Grainger College of Engineering, University of Illinois at Urbana\u2013Champaign. The BioTek Cytation 5 plate reader at the High Throughput Screening Facility, University of Illinois at Urbana\u2013Champaign, was mainly funded by the Office of the Director, National Institutes of Health, under award S10 OD025289. The icon of the influenza virus used in this study was obtained from BioRender.com.
This work was supported by the Strategic Research Initiatives Program at the Grainger College of Engineering, University of Illinois at Urbana-Champaign. The BioTek Cytation 5 plate reader at the High Throughput Screening Facility, University of Illinois at Urbana-Champaign, was mainly funded by the Office of the Director, National Institutes of Health, under award S10 OD025289. The icon of the influenza virus used in this study was obtained from BioRender.com.
PY - 2024/9/20
Y1 - 2024/9/20
N2 - Timely surveillance of airborne pathogens is essential to preventing the spread of infectious diseases and safeguard human health. Methods for sensitive, efficient, and cost-effective detection of airborne viruses are needed. With advances in synthetic biology, whole-cell biosensors have emerged as promising platforms for environmental monitoring and medical diagnostics. However, the current design paradigm of whole-cell biosensors is mostly based on intracellular detection of analytes that can transport across the cell membrane, which presents a critical challenge for viral pathogens and large biomolecules. To address this challenge, we developed a new type of whole-cell biosensor by expressing and displaying VHH-based quenchbody (Q-body) on the surface of the yeast Saccharomyces cerevisiae for simple one-step detection of influenza A (H1N1) virus. Seventeen VHH antibody fragments targeting the hemagglutinin protein H1N1-HA were displayed on the yeast cells and screened for the H1N1-HA binding affinity. The functionally displayed VHHs were selected to create surface-displayed Q-body biosensors. The surface-displayed Q-body exhibiting the highest quenching and dequenching efficiency was identified. The biosensor quantitatively detected H1N1-HA in a range from 0.5 to 16 μg/mL, with a half-maximal concentration of 2.60 μg/mL. The biosensor exhibited high specificity for H1N1-HA over other hemagglutinin proteins from various influenza A virus subtypes. Moreover, the biosensor succeeded in detecting the H1N1 virus at concentrations from 2.4 × 104 to 1.5 × 107 PFU/mL. The results from this study demonstrated a new whole-cell biosensor design that circumvents the need for transport of analytes into biosensor cells, enabling efficient detection of the target virus particles.
AB - Timely surveillance of airborne pathogens is essential to preventing the spread of infectious diseases and safeguard human health. Methods for sensitive, efficient, and cost-effective detection of airborne viruses are needed. With advances in synthetic biology, whole-cell biosensors have emerged as promising platforms for environmental monitoring and medical diagnostics. However, the current design paradigm of whole-cell biosensors is mostly based on intracellular detection of analytes that can transport across the cell membrane, which presents a critical challenge for viral pathogens and large biomolecules. To address this challenge, we developed a new type of whole-cell biosensor by expressing and displaying VHH-based quenchbody (Q-body) on the surface of the yeast Saccharomyces cerevisiae for simple one-step detection of influenza A (H1N1) virus. Seventeen VHH antibody fragments targeting the hemagglutinin protein H1N1-HA were displayed on the yeast cells and screened for the H1N1-HA binding affinity. The functionally displayed VHHs were selected to create surface-displayed Q-body biosensors. The surface-displayed Q-body exhibiting the highest quenching and dequenching efficiency was identified. The biosensor quantitatively detected H1N1-HA in a range from 0.5 to 16 μg/mL, with a half-maximal concentration of 2.60 μg/mL. The biosensor exhibited high specificity for H1N1-HA over other hemagglutinin proteins from various influenza A virus subtypes. Moreover, the biosensor succeeded in detecting the H1N1 virus at concentrations from 2.4 × 104 to 1.5 × 107 PFU/mL. The results from this study demonstrated a new whole-cell biosensor design that circumvents the need for transport of analytes into biosensor cells, enabling efficient detection of the target virus particles.
KW - influenza virus
KW - quenchbody (Q-body)
KW - virus detection
KW - whole-cell biosensor
KW - yeast surface display
UR - https://www.scopus.com/pages/publications/85204465969
UR - https://www.scopus.com/pages/publications/85204465969#tab=citedBy
U2 - 10.1021/acssynbio.4c00317
DO - 10.1021/acssynbio.4c00317
M3 - Article
C2 - 39256183
AN - SCOPUS:85204465969
SN - 2161-5063
VL - 13
SP - 2926
EP - 2937
JO - ACS synthetic biology
JF - ACS synthetic biology
IS - 9
ER -