A serological assay to detect SARS-CoV-2 seroconversion in humans

“…The ELISA assay was performed in a two-step process whereby a positivity screen was then followed by an endpoint titer determination following previously established protocols 43 . Briefly, 96-well EIA/RIA plates (Corning, Sigma) were coated with PBS or 0.1μg per well of either S-R/PP or S-R/x2 at 4°C overnight.…”

Section: Elisa Assaysmentioning

confidence: 99%

A thermostable, closed, SARS-CoV-2 spike protein trimer

Xiong

Ciazynska

et al. 2020

Preprint

169

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The spike (S) protein of SARS-CoV-2 mediates receptor binding and cell entry and is the dominant target of the immune system. S exhibits substantial conformational flexibility. It transitions from closed to open conformations to expose its receptor binding site, and subsequently from prefusion to postfusion conformations to mediate fusion of viral and cellular membranes. S protein derivatives are components of vaccine candidates and diagnostic assays, as well as tools for research into the biology and immunology of SARS-CoV-2. Here we have designed mutations in S which allow production of thermostable, crosslinked, S protein trimers that are trapped in the closed, pre-fusion, state. We have determined the structures of crosslinked and non-crosslinked proteins, identifying two distinct closed conformations of the S trimer. We demonstrate that the designed, thermostable, closed S trimer can be used in serological assays. This protein has potential applications as a reagent for serology, virology and as an immunogen.

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“…Slides were stained with DAPI (Sigma) and Enzyme-linked immunosorbent assay. ELISAs were performed as previously described 9 . In short, Triton X-100 and RNase A were added to serum samples at final concentrations of 0.5% and 0.5mg/ml respectively and incubated at room temperature (RT) for 3 hours before use to reduce risk from any potential virus in serum.…”

Section: Sars-covmentioning

confidence: 99%

“…The role of adaptive immunity and specifically antibody response to SARS-CoV-2 is particularly important in the development of safe and effective vaccines. To assess the capacity for B6J AAV-hACE2 mice to mount an antibody response to SARS-CoV-2 challenge, we quantified antispike protein IgG titers by ELISA 9,10 . We found that while control infected mice did not develop anti-spike antibodies, AAV-hACE2 B6/J mice mounted a significant antibody response between 4-and 7-DPI, which continued to increase at 14 DPI ( Fig.…”

mentioning

confidence: 99%

Mouse model of SARS-CoV-2 reveals inflammatory role of type I interferon signaling

Israelow

Song

Mao

et al. 2020

Preprint

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Severe Acute Respiratory Syndrome-Coronavirus 2 (SARS-Cov-2) has caused over 5,000,000 cases of Coronavirus disease (COVID-19) with significant fatality rate. 1-3 Due to the urgency of this global pandemic, numerous therapeutic and vaccine trials have begun without customary safety and efficacy studies. 4 Laboratory mice have been the stalwart of these types of studies; however, they do not support infection by SARS-CoV-2 due to the inability of its spike (S) protein to engage the mouse ortholog of its human entry receptor angiotensin-converting enzyme 2 (hACE2). While hACE2 transgenic mice support infection and pathogenesis, 5 these mice are currently limited in availability and are restricted to a single genetic background. Here we report the development of a mouse model of SARS-CoV-2 based on adeno associated virus (AAV)-mediated expression of hACE2. These mice support viral replication and antibody production and exhibit pathologic findings found in COVID-19 patients as well as non-human primate models. Moreover, we show that type I interferons are unable to control SARS-CoV2 replication and drive pathologic responses. Thus, the hACE2-AAV mouse model enables rapid deployment for in-depth analysis following robust SARS-CoV-2 infection with authentic patientderived virus in mice of diverse genetic backgrounds. This represents a much-needed platform for rapidly testing prophylactic and therapeutic strategies to combat COVID-19. Development of SARS-CoV-2 mouse model

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A serological assay to detect SARS-CoV-2 seroconversion in humans

Cited by 1,752 publications

References 26 publications

Serology assays to manage COVID-19

Serology assays to manage COVID-19

A thermostable, closed, SARS-CoV-2 spike protein trimer

Mouse model of SARS-CoV-2 reveals inflammatory role of type I interferon signaling

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