A cell-free high throughput assay for assessment of SARS-CoV-2 neutralizing antibodies

Autor: Jamil Yousef, Malin Jönsson, My Hedhammar, Sara Mravinacova, Anna Månberg, Peter Nilsson, Elisa Pin, Sebastian Havervall, Sophia Hober, Jonas Klingström, Wanda Christ, Cecilia Hellström, Charlotte Thålin, Hanna Tegel
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)
Population
Pseudoneutralization
Bioengineering
SPR
surface plasmon resonance
Cell free
AU
arbitrary unit
Antibodies
Viral
Neutralization
Antibodies
Serology
RBD
receptor binding domain
Immunity
Neutralization Tests
Microneutralization Assay
Humans
education
Molecular Biology
education.field_of_study
biology
ACE2
angiotensin converting enzyme 2
SARS-CoV-2
RPE
R-phycoerythrin conjugated
Cell-free
COVID-19
General Medicine
PNT
pseudoneutralization
WB
western blotting
HCW
healthcare workers
NC-C
C-terminal fragment of the nucleocapsid protein
Virology
Antibodies
Neutralizing
Full length Article
High-Throughput Screening Assays
HT
heat treatment
Spike Glycoprotein
Coronavirus
biology.protein
CV
coefficient of variance
Angiotensin-Converting Enzyme 2
Antibody
PFA
paraformaldehyde
Bead-based
Biotechnology
Zdroj: New Biotechnology
ISSN: 1876-4347
1871-6784
Popis: Highly accurate serological tests are key to assessing the prevalence of SARS-CoV-2 antibodies and the level of immunity in the population. This is important to predict the current and future status of the pandemic. With the recent emergence of new and more infectious SARS-CoV-2 variants, assays allowing for high throughput analysis of antibodies able to neutralize SARS-CoV-2 become even more important. Here, we report the development and validation of a robust, high throughput method, which enables the assessment of antibodies inhibiting the binding between the SARS-CoV-2 spike protein and angiotensin converting enzyme 2 (ACE2). The assay uses recombinantly produced spike-f and ACE2 and is performed in a bead array format, which allows analysis of up to 384 samples in parallel per instrument over seven hours, demanding only one hour of manual handling. The method is compared to a microneutralization assay utilising live SARS-CoV-2 and is shown to deliver highly correlating data. Further, a comparison with a serological method that measures all antibodies recognizing the spike protein shows that this type of assessment provides important insights into the neutralizing efficiency of the antibodies, especially for individuals with low antibody levels. This method can be an important and valuable tool for large-scale assessment of antibody-based neutralization, including neutralization of new spike variants that might emerge.
Databáze: OpenAIRE
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