A SARS‐CoV‐2 Neutralization Assay Using Single Molecule Arrays

Abstract

Coronavirus disease 2019 (COVID-19) manifests with high clinical variability and warrants sensitive and specific assays to analyze immune responses in infected and vaccinated individuals.U sing Single Molecule Arrays( Simoa), we developed an assay to assess antibody neutralization with high sensitivity and multiplexing capabilities based on antibodymediated blockage of the ACE2-spike interaction. The assay does not require live viruses or cells and can be performed in ab iosafety level 2l aboratory within two hours.W eu sed this assayt oa ssess neutralization and antibody levels in patients who died of COVID-19 and patients hospitalized for as hort period of time and showthat neutralization and antibody levels increase over time.Wealso adapted the assayfor SARS-CoV-2 variants and measured neutralization capacity in pre-pandemic healthy,C OVID-19 infected, and vaccinated individuals.T his assayi sh ighly adaptable for clinical applications,s uch as vaccine development and epidemiological studies.

Rapid quantitative electrochemical detection of SARS-CoV-2 antibodies in plasma and dried blood spot samples

“…4d). [18] We also performed an assay to observe if BSA in the antifouling coating and the blocking buffer could be replaced by recombinant human albumin (rHA) without compromising sensitivity and specificity of the assay. We explored rHA as it is blood and animal component-free recombinant human albumin excipient which is U.S. Food and Drug Administration (FDA) and (European Medicines Agency) EMA-approved excipient.…”

“…A time-point study of SARS-CoV-2 was also carried out to understand seroconversion better. Plasma samples were collected on 5 different days(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21) after the onset of symptoms of COVID-19 for each person. The study was performed with 3 different patients.…”

Ultrasensitive monitoring of SARS-CoV-2-specific antibody responses based on a digital approach reveals one week of IgG seroconversion

“…Single molecule array (Simoa), has shown unique advantages in clinical applications, especially in the detection of low abundance proteins or biomarkers due to its high sensitivity ( Preische et al, 2019 ). Compared with traditional ELISA tests, Simoa has also played a great role in the detection of antigens and antibodies ( Cai et al, 2021 ; Norman et al, 2020 ; Shan et al, 2021 ), neutralizing antibodies ( Gilboa et al, 2021 ) and vaccine monitoring ( Ogata et al, 2021 ) in body fluids during the COVID-19 pandemic, contributing greatly to the in-depth understanding of COVID-19 and the development of new detection and monitoring methods. However, most of the current seroconversion studies start from the onset of symptoms, while the number of days of infection before the onset of symptoms is unknown.…”

Integrated System for On-Site Rapid and Safe Screening of COVID-19

“…RT-PCR is the gold standard method for COVID-19 diagnosis but usually requires lengthy multi-step procession, professional operations, and costly infrastructure. , Serological assays including chemiluminescence immunoassay (CLIA) and immunochromatography assay (ICA) are on the basis of antigen-/antibody-specific binding and considered to be complements to RT-PCR. , Serological assays usually show high specificity (81.25–99.00%) and sensitivity (81.00–100.00%), , especially at >7 days after symptom onset . In addition, serological assays can help revealing the immune response in convalescents, vaccinated, or asymptomatic individuals . Nevertheless, serological methods are susceptible to endogenous disruptors or nonspecific antibodies, resulting in false-positive results .…”

“… 17 In addition, serological assays can help revealing the immune response in convalescents, vaccinated, or asymptomatic individuals. 18 Nevertheless, serological methods are susceptible to endogenous disruptors or nonspecific antibodies, resulting in false-positive results. 19 In addition, places such as airports may not be equipped with molecular diagnostic laboratories.…”