CPC G01N 33/54388 (2021.08) [G01N 33/56983 (2013.01); G01N 2333/165 (2013.01)] | 26 Claims |
1. A lateral flow device for the detection of antibodies to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in a sample of bodily fluid of an animal or human, the device comprising: a) a strip formed of a material enabling capillary flow of fluid along a portion of the strip; b) a sample pad located proximal to one end of the strip for receiving the sample of the bodily fluid, c) a conjugate pad located in the strip so that in operation the sample flows under capillary action through the strip from the sample pad to the conjugate pad and mobilizes a conjugate contained in the conjugate pad, the conjugate comprising a recombinant SARS-CoV-2 spike (S) protein antigen that has been conjugated to a detection agent, and d) a detection band comprising an Fc-binding protein immobilized within the strip along a band located perpendicular to the direction of flow of the sample along the strip so that when the mobilized spike protein conjugate in the sample contacts the Fc-binding protein in the detection band the presence of the antibodies to the SARS-CoV-2 virus in the sample is indicated by a visible color change, wherein the recombinant SARS-CoV-2 spike (S) protein antigen in c) is in a prefusion conformation and comprises a fragment of a wild-type SARS-CoV-2 protein having the amino acid sequence of SEQ ID NO: 1, wherein the fragment comprises the S1 and S2 domains and includes a double proline substitution at positions 986 and 987 of the wild-type protein and wherein the fragment further comprises a furin cleavage site “GSAS (SEQ ID NO: 5)” at positions 682 to 685 of the wild-type protein and a C-terminal T4 fibritin foldon motif “GYIPEAPRGDQAYVRKDGEWVLLSTFL” (SEQ ID NO: 2).
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