High-throughput quantitation of SARS-CoV-2 antibodies in a single-dilution homogeneous assay

Kainulainen, Markus H.; Bergeron, Éric; Chatterjee, Payel; Chapman, Asheley Poole; Lee, Joo Sang; Chida, Asiya Seema; Tang, Xiaoling; Wharton, Rebekah E.; Mercer, Kristina B.; Petway, Marla; Jenks, Harley M.; Flietstra, Timothy D.; Schuh, Amy J.; Satheshkumar, Panayampalli S.; Chaitram, Jasmine; Owen, S. Michele; McMullan, Laura K.; Flint, Mike; Finn, M. G.; Goldstein, Jason; Spiropoulou, Christina F.

doi:10.1038/s41598-021-91300-5

Cited by 14 publications

(17 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“… 12 − 14 Recent NanoBiT immunoassays with approximately 30 min time scales have reliably quantified several protein biomarkers and antibodies over concentrations spanning 4 orders of magnitude, with low pM sensitivity and up to a 1000-fold signal to background ratios. 14 , 15 Analytical performance approaches or exceeds laboratory EIAs, but with the ease and speed of LFTs. 14 , 15 …”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Rapid Quantification of C. difficile Glutamate Dehydrogenase and Toxin B (TcdB) with a NanoBiT Split-Luciferase Assay

et al. 2022

View full text Add to dashboard Cite

C. difficile infection (CDI) is a leading healthcare-associated infection with a high morbidity and mortality and is a financial burden. No current standalone point-of-care test (POCT) is sufficient for the identification of true CDI over a disease-free carriage of C. difficile , so one is urgently required to ensure timely, appropriate treatment. Here, two types of binding proteins, Affimers and nanobodies, targeting two C. difficile biomarkers, glutamate dehydrogenase (GDH) and toxin B (TcdB), are combined in NanoBiT (NanoLuc Binary Technology) split-luciferase assays. The assays were optimized and their performance controlling parameters were examined. The 44 fM limit of detection (LoD), 4–5 log range and 1300-fold signal gain of the TcdB assay in buffer is the best observed for a NanoBiT assay to date. In the stool sample matrix, the GDH and TcdB assay sensitivity (LoD = 4.5 and 2 pM, respectively) and time to result (32 min) are similar to a current, commercial lateral flow POCT, but the NanoBit assay has no wash steps, detects clinically relevant TcdB over TcdA, and is quantitative. Development of the assay into a POCT may drive sensitivity further and offer an urgently needed ultrasensitive TcdB test for the rapid diagnosis of true CDI. The NanoBiTBiP (NanoBiT with Binding Proteins) system offers advantages over NanoBiT assays with antibodies as binding elements in terms of ease of production and assay performance. We expect this methodology and approach to be generally applicable to other biomarkers.

show abstract

Section: Introductionmentioning

confidence: 99%

“… 14 , 15 Analytical performance approaches or exceeds laboratory EIAs, but with the ease and speed of LFTs. 14 , 15 …”

Section: Introductionmentioning

confidence: 99%

Rapid Quantification of C. difficile Glutamate Dehydrogenase and Toxin B (TcdB) with a NanoBiT Split-Luciferase Assay

et al. 2022

View full text Add to dashboard Cite

show abstract

“…This seemed a reasonable target based on recent accounts of highly sensitive split Nluc-based immunoassays using genetic fusions, where only a single reporter peptide is necessary for each molecule of affinity reagent. 7 − 9 , 11 , 25 , 26 …”

Section: Resultsmentioning

confidence: 99%

Simple, Rapid Chemical Labeling and Screening of Antibodies with Luminescent Peptides

et al. 2022

View full text Add to dashboard Cite

Sensitive and selective detection assays are essential for the accurate measurement of analytes in both clinical and research laboratories. Immunoassays that rely on nonoverlapping antibodies directed against the same target analyte (e.g., sandwich enzyme-linked immunosorbent assays (ELISAs)) are commonly used molecular detection technologies. Use of split enzyme reporters has simplified the workflow for these traditionally complex assays. However, identifying functional antibody pairs for a given target analyte can be cumbersome, as it generally involves generating and screening panels of antibodies conjugated to reporters. Accordingly, we sought a faster and easier reporter conjugation strategy to streamline antibody screening. We describe here the development of such a method that is based on an optimized ternary NanoLuc luciferase. This bioluminescence complementation system is comprised of a reagent-based thermally stable polypeptide (LgTrip) and two small peptide tags (β9 and β10) with lysine-reactive handles for direct conjugation onto antibodies. These reagents enable fast, single-step, wash-free antibody labeling and sensitive functional screening. Simplicity, speed, and utility of the one-pot labeling technology are demonstrated in screening antibody pairs for the analyte interleukin-4. The screen resulted in the rapid development of a sensitive homogeneous immunoassay for this clinically relevant cytokine.

show abstract

“…Blood specimens ( n = 6) were collected in 5 ml vacutainer serum separator tubes from the cephalic vein of both dogs at three time points. Serological response against SARS‐CoV‐2 was determined using a species‐independent assay detecting antibody binding to the receptor‐binding domain (RBD) of the viral spike (S) protein (Kainulainen et al., 2021 ).…”

Section: Methodsmentioning

confidence: 99%

Transmission of SARS‐CoV‐2 Delta variant (B.1.617.2) from a fully vaccinated human to a canine in Georgia, July 2021

Wendling

Carpenter

Liew

et al. 2022

Zoonoses and Public Health

Self Cite

View full text Add to dashboard Cite

SARS‐CoV‐2 infection has been described in a wide range of species, including domestic animals such as dogs and cats. Illness in dogs is usually self‐limiting, and further diagnostics may not be pursued if clinical signs resolve or they respond to empirical treatment. As new variants emerge, the clinical presentation and role in transmission may vary in animals. This report highlights different clinical presentations and immunological responses in two SARS‐CoV‐2 Delta‐variant‐positive dogs with similar exposure to the same fully vaccinated human with a SARS‐CoV‐2 infection and emphasizes the need for active surveillance and additional One Health research on SARS‐CoV‐2 variant infections in companion animals and other species.

show abstract

High-throughput quantitation of SARS-CoV-2 antibodies in a single-dilution homogeneous assay

Cited by 14 publications

References 35 publications

Rapid Quantification of C. difficile Glutamate Dehydrogenase and Toxin B (TcdB) with a NanoBiT Split-Luciferase Assay

Rapid Quantification of C. difficile Glutamate Dehydrogenase and Toxin B (TcdB) with a NanoBiT Split-Luciferase Assay

Simple, Rapid Chemical Labeling and Screening of Antibodies with Luminescent Peptides

Transmission of SARS‐CoV‐2 Delta variant (B.1.617.2) from a fully vaccinated human to a canine in Georgia, July 2021

Contact Info

Product

Resources

About