Methods, Compositions, and Systems for Detecting Coronavirus Neutralizing Antibodies
AssigneeLaboratory Corporation of America Holdings
InventorsChristos J. Petropoulos, Mary T. Wrin, Danielle DiTirro
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Abstract
The present disclosure relates to methods, compositions, and systems for detecting whether a subject exposed to a coronavirus has developed a neutralizing antibody response. Also disclosed are methods for determining whether a patient infected by a coronavirus is likely to respond to treatment with an antibody preparation. Also disclosed are methods for detecting the level of neutralizing antibody response in a sample of serum from a subject exposed to a coronavirus or to a coronavirus vaccine.
Claims20
A method for detecting whether a subject exposed to a coronavirus has developed a neutralizing antibody response, comprising:
- (a) transfecting into a plurality of first cells
- i) a nucleic acid encoding a coronavirus spike protein, and
- ii) a viral expression vector which comprises an indicator nucleic acid which produces a detectable signal;
- (b) incubating the first cells under conditions such that the first cells produce viral particles comprising the coronavirus spike protein;
- (c) contacting the viral particles of step (b) with a second cell in the presence or absence of a sample comprising an antibody from the subject, wherein the second cell expresses a cell surface receptor to which the coronavirus binds;
- (d) measuring the amount of the detectable signal produced by the second cell in the presence or absence of the sample;
- and
- (e) comparing the amount of signal measured in step (d) in the presence of the sample with the amount of signal produced in step (d) in the absence of the sample, wherein a reduced amount of signal measured in the presence of the sample indicates that the subject has developed a neutralizing antibody response capable of reducing infection.
A method for evaluating the effect of a mutation in the spike protein on susceptibility to anti-SARS CoV-2 antibodies, comprising:
- (a) transfecting into a first portion of a plurality of first cells:
- i) a nucleic acid encoding a control coronavirus spike protein, and
- ii) a viral expression vector which comprises an indicator nucleic acid which produces a detectable signal;
- (b) transfecting into a second portion of a plurality of first cells:
- i) a nucleic acid encoding a coronavirus spike protein that comprises a mutation;
- and
- ii) a viral expression vector which comprises an indicator nucleic acid which produces a detectable signal;
- (c) incubating the first and second portion of first cells separately under conditions such that the first portion cells produce first viral particles comprising the control coronavirus spike protein, and the second portion cells produce second viral particles comprising the coronavirus spike protein having the mutation;
- (d) contacting the first viral particles of step (c) with a first portion of a plurality of second cells in the presence of a sample comprising an antibody that binds SARS CoV-2, wherein the second cell expresses a cell surface receptor to which the coronavirus binds;
- (e) contacting the second viral particles of step (c) with a second portion of a plurality of second cells in the presence of a sample comprising an antibody that binds SARS CoV-2, wherein the second cell expresses a cell surface receptor to which the coronavirus binds;
- and
- (f) measuring the amount of the detectable signal produced by the second cell in steps (d) and (e), wherein a reduced amount of signal produced by the second cell in step (d) as compared to the signal produced by the second cell in step (e) indicates that the mutation in the spike protein confers a reduced susceptibility of the viral particles to the anti-SARS CoV-2 antibody.
A composition for the detection of a compound that can modulate SARS infectivity, comprising a cell that has been genetically modified to be either a producer cell or a target cell.
A method for detecting the level of neutralizing antibody response in a sample from a subject exposed to a coronavirus vaccine, comprising:
- (a) transfecting into a plurality of first cells:
- i) a nucleic acid encoding a coronavirus spike protein, and
- ii) a viral expression vector which comprises an indicator nucleic acid which produces a detectable signal;
- (b) incubating the first cells under conditions such that the first cells produce viral particles comprising the coronavirus spike protein;
- (c) contacting the viral particles of step (b) with a second cell in the presence of a sample from the subject obtained before the subject was exposed to a coronavirus vaccine or a sample from the subject obtained after the subject was exposed to a coronavirus vaccine, wherein the second cell expresses a cell surface receptor to which the coronavirus binds;
- (d) measuring the amount of the detectable signal produced by the second cell in the presence of the sample obtained before the subject was exposed to the coronavirus vaccine;
- (e) measuring the amount of the detectable signal produced by the second cell in the presence of the sample obtained after the subject was exposed to the coronavirus vaccine;
- and
- (f) comparing the amount of signal measured in step (d) with the amount of signal produced in step (e), and determining the level of a neutralizing antibody response in the subject based on the difference in the amount of signal measured in step (d) with the amount of signal produced in step (e).
Dependencies are inferred from the claim text.