SERS-based assay for multiplexed detection of cross-reactivity and persistence of antibodies against the spike of the native, P.1 and B.1.617.2 SARS-CoV-2 in non-hospitalised adults

Chisanga, Malama; Stuible, Matthew; Gervais, Christian; L’Abbé, Denis; Cass, Brian; Bisson, Louis; Pelletier, Alex; Lord‐Dufour, Simon; Durocher, Yves; Boudreau, Denis; Trottier, Sylvie; Pelletier, Joelle N.; Masson, Jean-François

doi:10.1039/d2sd00073c

Cited by 5 publications

(7 citation statements)

References 67 publications

(110 reference statements)

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“…The gold coating and the peptide self‐assembled monolayer (SAM) on the SPR detection prisms were optimized as previously described 37–44 . Briefly, bare prisms were coated with 2 nm of chromium and 50 nm of gold with a Cressington coating system.…”

Section: Methodsmentioning

confidence: 99%

“…The gold coating and the peptide self-assembled monolayer (SAM) on the SPR detection prisms were optimized as previously described. [37][38][39][40][41][42][43][44] Briefly, bare prisms were coated with 2 nm of chromium and 50 nm of gold with a Cressington coating system. All prisms were then coated overnight with a peptide SAM (3MPA-LHD-LHD-COOH) solution of 100 μg/mL in DMF (N-N-dimethylformamide).…”

Section: Preparation Of the Spr Gold Prismsmentioning

confidence: 99%

“…The initial steps were similar to those previously described for other analytes. [37][38][39][40][41][42][43][44] Briefly, a 1:1 ratio of 400 mM aqueous EDC (N-[3-dimethylaminopropyl]-N 0ethylcarbodiimide hydrochloride) and 100 mM aqueous NHS (N-hydroxysuccinimide) was injected for 2 min (120 s) into each channel to activate the SAM. An aqueous solution (500 μL) of 10 mM sodium acetate (pH 4.5) was then injected for 30 s to optimize the environment and wash off the previous reagent.…”

Section: Construction Of the Iga Biosensormentioning

confidence: 99%

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A portable surface plasmon resonance (SPR) sensor for the detection of immunoglobulin A in plasma

Dubois,

Ducas,

Laforce‐Lavoie

et al. 2024

Transfusion

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BackgroundA life‐threatening anaphylactic shock can occur if a patient with undiagnosed immunoglobulin A (IgA) deficiency (i.e., IgA levels <500 ng/mL) receives IgA‐containing blood, hence the need for a rapid, point‐of‐care (POC) method for IgA deficiency screening. Enzyme‐linked immunosorbent assay (ELISA) is routinely used to detect IgA, but this method requires trained specialists and ≥24 h to obtain a result. We developed a surface plasmon resonance (SPR)‐based protocol to identify IgA‐deficient patients or donors within 1 h.Materials and MethodsThe SPR sensor relies on the detection of IgAs captured by primary antibodies adsorbed on the SPR chip and quantified with secondary antibodies. The sensor was calibrated from 0 to 2000 ng/mL in buffer, IgA‐depleted human serum, and plasma samples from IgA‐deficient individuals. A critical concentration of 500 ng/mL was set for IgA deficiency. The optimized sensor was then tested on eight plasma samples with known IgA status (determined by ELISA), including five with IgA deficiency and three with normal IgA levels.ResultsThe limit of detection was estimated at 30 ng/mL in buffer and 400 ng/mL in diluted plasma. The results obtained fully agreed with ELISA among the eight plasma samples tested. The protocol distinguished IgA‐deficient from normal samples, even for samples with an IgA concentration closer to critical concentration.DiscussionIn conclusion, we developed a reliable POC assay for the quantification of IgA in plasma. This test may permit POC testing at blood drives and centralized centers to maintain reserves of IgA‐deficient blood and in‐hospital testing of blood recipients.

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Section: Methodsmentioning

confidence: 99%

Section: Preparation Of the Spr Gold Prismsmentioning

confidence: 99%

Section: Construction Of the Iga Biosensormentioning

confidence: 99%

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A portable surface plasmon resonance (SPR) sensor for the detection of immunoglobulin A in plasma

Dubois,

Ducas,

Laforce‐Lavoie

et al. 2024

Transfusion

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“…Participants with mild COVID-19 were randomly selected from the same cohort described previously. , All the volunteers were 18 years or older and had received a PCR-confirmed positive test for SARS-CoV-2 between 2 and 3 weeks prior to enrolment. Serum sampling was done at 4 and 16 weeks post-diagnosis.…”

Section: Experimental Methodsmentioning

confidence: 99%

Label-Free SERS for Rapid Differentiation of SARS-CoV-2-Induced Serum Metabolic Profiles in Non-Hospitalized Adults

et al. 2023

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COVID-19 represents a multi-system infectious disease with broad-spectrum manifestations, including changes in host metabolic processes connected to the disease pathogenesis. Understanding biochemical dysregulation patterns as a consequence of COVID-19 illness promises to be crucial for tracking disease course and clinical outcomes. Surface-enhanced Raman scattering (SERS) has attracted considerable interest in biomedical diagnostics for the sensitive detection of intrinsic profiles of unique fingerprints of serum biomolecules indicative of SARS-CoV-2 infection in a label-free format. Here, we applied label-free SERS and chemometrics for rapid interrogation of temporal metabolic dynamics in longitudinal sera of mildly infected non-hospitalized patients (n = 22), at 4 and 16 weeks post PCR-positive diagnosis, and compared them with negative controls (n = 8). SERS spectral markers revealed distinct metabolic profiles in patient sera that significantly deviated from the healthy metabolic state at the two sampling time intervals. Multivariate and univariate analyses of the spectral data identified abundance dynamics in amino acids, lipids, and protein vibrations as the key spectral features underlying the metabolic differences detected in convalescent samples and perhaps associated with patient recovery progression. A validation study performed using spontaneous Raman spectroscopy yielded spectral data results that corroborated SERS spectral findings and confirmed the detected disease-specific molecular phenotypes in clinical samples. Label-free SERS promises to be a valuable analytical technique for rapid screening of the metabolic phenotype induced by SARS-CoV-2 infection to allow appropriate healthcare intervention.

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“…In response to the COVID-19 pandemic, an important focus of our research has been the production of recombinant SARS-CoV-2 spike proteins in CHO cells. These proteins, specifically soluble, trimeric, prefusion-stabilized spike ectodomain constructs (Stuible et al, 2021), have been investigated as candidate vaccine antigens and also as reagents for serosurveillance studies (Akache et al, 2021(Akache et al, , 2022Chisanga et al, 2022;Colwill et al, 2022;Rudi et al, 2022;Stark et al, 2022;Stocks et al, 2021). With the frequent emergence of new variants, we have had the opportunity to generate dozens of spike-expressing CHO pools over the last 2 years, which has allowed us to appreciate better the performance and robustness of this expression platform.…”

Section: Introductionmentioning

confidence: 99%

A CHO stable pool production platform for rapid clinical development of trimeric SARS‐CoV‐2 spike subunit vaccine antigens

Joubert

Stuible

Lord‐Dufour

et al. 2023

Biotech & Bioengineering

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Protein expression from stably transfected Chinese hamster ovary (CHO) clones is an established but time‐consuming method for manufacturing therapeutic recombinant proteins. The use of faster, alternative approaches, such as non‐clonal stable pools, has been restricted due to lower productivity and longstanding regulatory guidelines. Recently, the performance of stable pools has improved dramatically, making them a viable option for quickly producing drug substance for GLP‐toxicology and early‐phase clinical trials in scenarios such as pandemics that demand rapid production timelines. Compared to stable CHO clones which can take several months to generate and characterize, stable pool development can be completed in only a few weeks. Here, we compared the productivity and product quality of trimeric SARS‐CoV‐2 spike protein ectodomains produced from stable CHO pools or clones. Using a set of biophysical and biochemical assays we show that product quality is very similar and that CHO pools demonstrate sufficient productivity to generate vaccine candidates for early clinical trials. Based on these data, we propose that regulatory guidelines should be updated to permit production of early clinical trial material from CHO pools to enable more rapid and cost‐effective clinical evaluation of potentially life‐saving vaccines.

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SERS-based assay for multiplexed detection of cross-reactivity and persistence of antibodies against the spike of the native, P.1 and B.1.617.2 SARS-CoV-2 in non-hospitalised adults

Abstract: Monitoring antibody response to SARS-CoV-2 is critical for assessing the humoral response, especially important considering the emergence of multiple SARS-CoV-2 variants of concern (VOCs). Herein, we developed microfluidics-integrated multiplexed SERS...

Cited by 5 publications

References 67 publications

A portable surface plasmon resonance (SPR) sensor for the detection of immunoglobulin A in plasma

A portable surface plasmon resonance (SPR) sensor for the detection of immunoglobulin A in plasma

Label-Free SERS for Rapid Differentiation of SARS-CoV-2-Induced Serum Metabolic Profiles in Non-Hospitalized Adults

A CHO stable pool production platform for rapid clinical development of trimeric SARS‐CoV‐2 spike subunit vaccine antigens

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