Engineering luminescent biosensors for point-of-care SARS-CoV-2 antibody detection

Elledge, Susanna K.; Zhou, Xin; Byrnes, James R.; Martinko, Alexander J.; Lui, Irene; Pance, Katarina; Lim, Shion A.; Glasgow, Jeff E.; Glasgow, Anum; Turcios, Keirstinne; Iyer, Nikita S.; Torres, Leonel; Peluso, Michael J.; Henrich, Timothy J.; Wang, Taia T.; Tato, Cristina M.; Leung, Kevin; Greenhouse, Bryan; Wells, James A.

doi:10.1101/2020.08.17.20176925

Cited by 21 publications

(32 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Supplementary Table 1 describes each of the assays performed as part of this analysis, including their sensitivity and specificity (as reported by the manufacturers for commercial assays, or by validation testing for non-commercial research assays). Supplementary Tables 2 and 3), 19 the split luciferase assay (total Ig) targeting N and S performed in the Wells laboratory, 20 and the Luminex assay (IgG) targeting N (one full-length and one fragment), S, and RBD performed in the Greenhouse laboratory.…”

Section: Antibody Assaysmentioning

confidence: 99%

SARS-CoV-2 antibody magnitude and detectability are driven by disease severity, timing, and assay

Peluso

Takahashi

Hakim

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Serosurveillance studies are critical for estimating SARS-CoV-2 transmission and immunity, but interpretation of results is currently limited by poorly defined variability in the performance of antibody assays to detect seroreactivity over time in individuals with different clinical presentations. We measured longitudinal antibody responses to SARS-CoV-2 in plasma samples from a diverse cohort of 128 individuals over 160 days using 14 binding and neutralization assays. For all assays, we found a consistent and strong effect of disease severity on antibody magnitude, with fever, cough, hospitalization, and oxygen requirement explaining much of this variation. We found that binding assays measuring responses to spike protein had consistently higher correlation with neutralization than those measuring responses to nucleocapsid, regardless of assay format and sample timing. However, assays varied substantially with respect to sensitivity during early convalescence and in time to seroreversion. Variations in sensitivity and durability were particularly dramatic for individuals with mild infection, who had consistently lower antibody titers and represent the majority of the infected population, with sensitivities often differing substantially from reported test characteristics (e.g., amongst commercial assays, sensitivity at 6 months ranged from 33% for ARCHITECT IgG to 98% for VITROS Total Ig). Thus, the ability to detect previous infection by SARS-CoV-2 is highly dependent on the severity of the initial infection, timing relative to infection, and the assay used. These findings have important implications for the design and interpretation of SARS-CoV-2 serosurveillance studies.

show abstract

Section: Antibody Assaysmentioning

confidence: 99%

SARS-CoV-2 antibody magnitude and detectability are driven by disease severity, timing, and assay

Peluso

Takahashi

Hakim

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…20 ). A similar approach based on intensiometric detection of split NanoLuc complementation was recently published as a preprint 55 , but the ratiometric detection enabled by the RAPPID assay is preferred for quantitative measurements, which are important when studying the time-dependence of the antibody response and the correlation between antibody titers and immunological protection 56,57 . Collectively, these results illustrate that the RAPPID platform is highly versatile and can be reengineered to suit a variety of applications related to antibody detection, including quantitative measurements of antibodies that bind discontinuous epitopes, therapeutic antibodies and their ADAs, and anti-SARS-CoV-2 antibodies.…”

Section: Resultsmentioning

confidence: 99%

“…We envision that the use of a calibrator luciferase can be applied as a general strategy to translate intensiometric luminescent assays into robust ratiometric assays with a time-independent and quantitative assay output. As such, other recently developed intensiometric sensors based on split luciferase complementation could benefit from a calibrator luciferase to further enhance their potential in a clinically relevant environment 21,55 . Furthermore, the concept can be extended by using different acceptor domains, such as quantum dots, organic fluorophores or fluorescent protein domains, with a larger spectral separation from the blue light-emitting NanoLuc.…”

Section: Conclusion and Discussionmentioning

confidence: 99%

RAPPID: a platform of ratiometric bioluminescent sensors for homogeneous immunoassays

Rosier

Aalen

et al. 2020

Preprint

View full text Add to dashboard Cite

Heterogeneous immunoassays such as ELISA have become indispensable in modern bioanalysis, yet translation into easy-to-use point-of-care assays is hindered by their dependence on external calibration and multiple washing and incubation steps. Here, we introduce RAPPID (Ratiometric Plug-and-Play Immunodiagnostics), a "mix-and-measure" homogeneous immunoassay platform that combines highly specific antibody-based detection with a ratiometric bioluminescent readout that can be detected using a basic digital camera. The concept entails analyte-induced complementation of split NanoLuc luciferase fragments, photoconjugated to an antibody sandwich pair via protein G adapters. We also introduce the use of a calibrator luciferase that provides a robust ratiometric signal, allowing direct in-sample calibration and quantitative measurements in complex media such as blood plasma. We developed RAPPID sensors that allow low-picomolar detection of several protein biomarkers, anti-drug antibodies, therapeutic antibodies, and both SARS-CoV-2 spike protein and anti-SARS-CoV-2 antibodies. RAPPID combines ratiometric bioluminescent detection with antibody-based target recognition into an easy-to-implement standardized workflow, and therefore represents an attractive, fast, and low-cost alternative to traditional immunoassays, both in an academic setting and in clinical laboratories for point-of-care applications.

show abstract

“…Photoluminescence is a very sensitive technique that can be applied in the design of various affinity biosensors for the determination of pathological cells [ 128 ] and virus-induced diseases [ 129 , 130 , 131 ]. Some researchers designed a split luciferase (spLUC) based antibody test that is showing itself as simple (not need ‘washing’, two-stage of reagent addition, rapid (less than 5 min), reliable (≥98%), low-volume specimen (1 µL for 1 reaction), inexpensive and solution-based quantitative approach to identify antibodies against S- and N- proteins of SARS-CoV-2 [ 132 ]. The biosensor was designed by merging small BiT (SmBiT) and large BiT (LgBiT) fragments [ 133 ] of Nanoluciferase (NanoLuc) to viral protein antigens.…”

Section: Affinity Biosensors For Covid-19 Diagnosismentioning

confidence: 99%

“…The ordinary differential equation modeling was executed to describe the ratio between signal intensity and immunoglobulin concentration and it was shown that there was a linear correlation between the specific antibody concentration and luciferase signal. The sensor showed sensitivity of 89% towards S-protein and 98% towards N-protein [ 132 ].…”

Section: Affinity Biosensors For Covid-19 Diagnosismentioning

confidence: 99%

Affinity Sensors for the Diagnosis of COVID-19

et al. 2021

View full text Add to dashboard Cite

The coronavirus disease 2019 (COVID-19) outbreak caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was proclaimed a global pandemic in March 2020. Reducing the dissemination rate, in particular by tracking the infected people and their contacts, is the main instrument against infection spreading. Therefore, the creation and implementation of fast, reliable and responsive methods suitable for the diagnosis of COVID-19 are required. These needs can be fulfilled using affinity sensors, which differ in applied detection methods and markers that are generating analytical signals. Recently, nucleic acid hybridization, antigen-antibody interaction, and change of reactive oxygen species (ROS) level are mostly used for the generation of analytical signals, which can be accurately measured by electrochemical, optical, surface plasmon resonance, field-effect transistors, and some other methods and transducers. Electrochemical biosensors are the most consistent with the general trend towards, acceleration, and simplification of the bioanalytical process. These biosensors mostly are based on the determination of antigen-antibody interaction and are robust, sensitive, accurate, and sometimes enable label-free detection of an analyte. Along with the specification of biosensors, we also provide a brief overview of generally used testing techniques, and the description of the structure, life cycle and immune host response to SARS-CoV-2, and some deeper details of analytical signal detection principles.

show abstract

Engineering luminescent biosensors for point-of-care SARS-CoV-2 antibody detection

Cited by 21 publications

References 60 publications

SARS-CoV-2 antibody magnitude and detectability are driven by disease severity, timing, and assay

SARS-CoV-2 antibody magnitude and detectability are driven by disease severity, timing, and assay

RAPPID: a platform of ratiometric bioluminescent sensors for homogeneous immunoassays

Affinity Sensors for the Diagnosis of COVID-19

Contact Info

Product

Resources

About