US 12,169,200 B2
Lateral flow device for detecting SARS-CoV-2 antibodies in human and animal samples
Joshua T. Lizer, Augusta, MI (US); Jason J. Workman, Wayland, MI (US); James P. Gillies, Allegan, MI (US); Eric T. Baima, Kalamazoo, MI (US); and Rajesh K. Mehra, Hayward, CA (US)
Assigned to Zoetis Services LLC, Parsippany, NJ (US)
Filed by Zoetis Services LLC, Parsippany, NJ (US)
Filed on Jul. 30, 2021, as Appl. No. 17/389,473.
Claims priority of provisional application 63/064,225, filed on Aug. 11, 2020.
Prior Publication US 2022/0050102 A1, Feb. 17, 2022
Int. Cl. G01N 33/543 (2006.01); G01N 33/569 (2006.01)
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).