Coronavirus proteins and antigens

Abstract

Disclosed herein are embodiments of a method for collecting, extracting or eluting proteins and antigens from cells infected with coronavirus. The coronavirus may be a porcine coronavirus, such as porcine epidemic diarrhea virus (PEDV) or porcine delta coronavirus (PDCoV). Also disclosed are embodiments of a composition comprising the coronavirus proteins and antigens, and embodiments of a method of using such a composition. Applications for the composition include, but are not limited to, use in the preparation of antibodies against the proteins and antigens, use as reference markers for coronavirus proteins, and/or use in an immunogenic composition, such as in a vaccine composition.

We claim: 
     
         1 . A method, comprising:
 isolating coronavirus-infected cells away from cell-free coronavirus viral particles in a population of cells in a culture medium; and   extracting or eluting coronavirus proteins and antigens from the isolated coronavirus-infected cells with a detergent-containing solution to form a solution comprising detergent and isolated coronavirus proteins and antigens.   
     
     
         2 . The method of  claim 1 , wherein the coronavirus is a porcine coronavirus. 
     
     
         3 . The method of  claim 2 , wherein the porcine coronavirus is transmissible gastroenteritis virus, porcine respiratory coronavirus, porcine epidemic diarrhea virus (PEDV), porcine hemagglutinating encephalomyelitis virus, porcine deltacoronavirus (PDCoV), or a combination thereof. 
     
     
         4 . The method of  claim 1 , further comprising allowing the population of cells to incubate for a time period sufficient to result in one or more replicated coronavirus viral particles being released into the culture medium. 
     
     
         5 . The method of  claim 1 , wherein the detergent-containing solution comprises a non-ionic detergent. 
     
     
         6 . The method of  claim 5 , wherein the non-ionic detergent is poly(ethylene glycol) p-isooctyl-phenyl ether, octylphenoxypolyethoxyethanol (Nonidet P-40), 4-(1,1,3,3-tetramethylbutyl)phenyl-polyethylene glycol (Triton X-100), or a combination thereof. 
     
     
         7 . The method of  claim 5 , wherein the detergent-containing solution comprises a detergent at a concentration of greater than zero to 5% in solution. 
     
     
         8 . The method of  claim 5 , wherein the detergent-containing solution comprises a chelating agent at a concentration of greater than zero to 0.5M in solution. 
     
     
         9 . The method of  claim 1 , wherein extracting or eluting coronavirus proteins and antigens comprises extracting or eluting coronavirus proteins and antigens for about 0.1 to about 15 hours. 
     
     
         10 . The method of  claim 9 , wherein extracting or eluting coronavirus proteins and antigens is performed at a temperature of from 0° C. to 25° C. 
     
     
         11 . The method of  claim 1 , wherein the proteins and antigens include coronavirus envelope proteins. 
     
     
         12 . The method of  claim 1 , wherein the solution of isolated coronavirus proteins and antigens has a concentration of coronavirus proteins and antigens greater than a concentration of coronavirus proteins and antigens in the culture medium that contained the population of infected cells. 
     
     
         13 . The method of  claim 1 , further comprising adding an adjuvant to the solution of isolated coronavirus proteins and antigens. 
     
     
         14 . The method of  claim 1 , further comprising adding an excipient to the solution of isolated coronavirus proteins and antigens. 
     
     
         15 . A composition comprising the isolated coronavirus proteins and antigens from  claim 1 , and an adjuvant. 
     
     
         16 . The composition of  claim 15 , comprising PEDV proteins and antigens, cell fragments, the detergent and an adjuvant. 
     
     
         17 . The composition of  claim 15 , comprising PDCoV proteins and antigens, cell fragments, the detergent and an adjuvant. 
     
     
         18 . A method of vaccinating a subject, the method comprising administering to the subject a composition comprising the solution of isolated coronavirus proteins and antigens from  claim 1 , 
     
     
         19 . The method of  claim 18 , wherein the subject is a porcine subject. 
     
     
         20 . A method of generating an immune response in a porcine subject, the method comprising administering to the subject the composition of  claim 15 .