TY - JOUR
T1 - Synthetic nanobody-functionalized nanoparticles for accelerated development of rapid, accessible detection of viral antigens
AU - Chen, Xiahui
AU - Kang, Shoukai
AU - Ikbal, Md Ashif
AU - Zhao, Zhi
AU - Pan, Yuxin
AU - Zuo, Jiawei
AU - Gu, Liangcai
AU - Wang, Chao
N1 - Publisher Copyright:
© 2022 Elsevier B.V.
PY - 2022/4/15
Y1 - 2022/4/15
N2 - Successful control of emerging infectious diseases requires accelerated development of fast, affordable, and accessible assays for wide implementation at a high frequency. This paper presents a design for an in-solution assay pipeline, featuring nanobody-functionalized nanoparticles for rapid, electronic detection (Nano2RED) of Ebola and COVID-19 antigens. Synthetic nanobody binders with high affinity, specificity, and stability are selected from a combinatorial library and site-specifically conjugated to gold nanoparticles (AuNPs). Without requiring any fluorescent labelling, washing, or enzymatic amplification, these multivalent AuNP sensors reliably transduce antigen binding signals upon mixing into physical AuNP aggregation and sedimentation processes, displaying antigen-dependent optical extinction readily detectable by spectrometry or portable electronic circuitry. With Ebola virus secreted glycoprotein (sGP) and a SARS-CoV-2 spike protein receptor binding domain (RBD) as targets, Nano2RED showed a high sensitivity (the limit of detection of ∼10 pg /mL, or 0.13 pM for sGP and ∼40 pg/mL, or ∼1.3 pM for RBD in diluted human serum), a high specificity, a large dynamic range (∼7 logs),and fast readout within minutes. The rapid detection, low material cost (estimated <$0.01 per test), inexpensive and portable readout system (estimated <$5), and digital data output, make Nano2RED a particularly accessible assay in screening of patient samples towards successful control of infectious diseases.
AB - Successful control of emerging infectious diseases requires accelerated development of fast, affordable, and accessible assays for wide implementation at a high frequency. This paper presents a design for an in-solution assay pipeline, featuring nanobody-functionalized nanoparticles for rapid, electronic detection (Nano2RED) of Ebola and COVID-19 antigens. Synthetic nanobody binders with high affinity, specificity, and stability are selected from a combinatorial library and site-specifically conjugated to gold nanoparticles (AuNPs). Without requiring any fluorescent labelling, washing, or enzymatic amplification, these multivalent AuNP sensors reliably transduce antigen binding signals upon mixing into physical AuNP aggregation and sedimentation processes, displaying antigen-dependent optical extinction readily detectable by spectrometry or portable electronic circuitry. With Ebola virus secreted glycoprotein (sGP) and a SARS-CoV-2 spike protein receptor binding domain (RBD) as targets, Nano2RED showed a high sensitivity (the limit of detection of ∼10 pg /mL, or 0.13 pM for sGP and ∼40 pg/mL, or ∼1.3 pM for RBD in diluted human serum), a high specificity, a large dynamic range (∼7 logs),and fast readout within minutes. The rapid detection, low material cost (estimated <$0.01 per test), inexpensive and portable readout system (estimated <$5), and digital data output, make Nano2RED a particularly accessible assay in screening of patient samples towards successful control of infectious diseases.
KW - COVID-19
KW - Co-binding nanobodies
KW - Ebola virus
KW - Infectious diseases
KW - Metal nanoparticles
KW - Rapid electronic detection
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U2 - 10.1016/j.bios.2022.113971
DO - 10.1016/j.bios.2022.113971
M3 - Article
C2 - 35051851
AN - SCOPUS:85122928184
SN - 0956-5663
VL - 202
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
M1 - 113971
ER -