Method of purifying virus-like particles

Abstract

A method of producing purified FMDV VLPs, comprising contacting cells containing FMDV VLPs with a lysis buffer and allowing the cells to lyse, the lysis buffer comprising 10-20 mM Tris-HCl, 150-200 mM NaCl, 3 mM MgCl2, and 1% Triton X-100, wherein the lysis buffer does not contain EDTA; centrifuging a solution; and removing a supernatant from the solution, the supernatant containing the purified FMDV VLPs.

1 . A method of producing purified VLPs, comprising:
 a) contacting cells containing VLPs with a lysis buffer and allowing the cells to lyse, the lysis buffer comprising:
 10-20 mM Tris-HCl, 
 150-200 mM NaCl, 
 3 mM MgCl 2 , 
 and 1% Triton X-100, 
 wherein the lysis buffer does not contain EDTA; 
   b) centrifuging a solution of step (a); and   c) removing a supernatant from the solution of step (b), the supernatant containing the purified VLPs.   
     
     
         2 . The method of  claim 1 , further comprising centrifuging the supernatant through a cesium chloride and/or sucrose gradient. 
     
     
         3 . The method of  claim 1 , wherein the cells are eukaryotic cells. 
     
     
         4 . The method of  claim 1 , wherein the cells are mammalian cells. 
     
     
         5 . The method of  claim 1 , wherein the cells are insect cells. 
     
     
         6 . The method of  claim 1 , wherein the cells are CHO-K7 or HEK293-T cells. 
     
     
         7 . The method of  claim 1 , wherein the VLPs is selected from Swine Vesicular Disease Virus, Senecavirus A, Porcine Teschovirus, Bovine rhinitis A virus, Bovine rhinitis B virus, Equine rhinitis A virus, or Foot-and-mouth disease virus (FMDV). 
     
     
         8 . A method of inoculating an animal to induce an immune response to viral disease, comprising: a) providing purified VLPs produced according to the method of  claim 1 ; and b) inoculating the animal with an effective amount of the VLPs. 
     
     
         9 . The method of  claim 8 , wherein the viral disease is selected from Swine Vesicular Disease Virus, Senecavirus A, Porcine Teschovirus, Bovine rhinitis A virus, Bovine rhinitis B virus, Equine rhinitis A virus, or Foot-and-mouth disease virus (FMDV) 
     
     
         10 . The method of  claim 8 , wherein the animal is a mammal. 
     
     
         11 . The method of  claim 8 , wherein the animal is a pig, cow, goat, horse or sheep. 
     
     
         12 . The method of  claim 11 , wherein the animal is a cow. 
     
     
         13 . The method of  claim 11 , wherein the animal is a pig. 
     
     
         14 . The method of  claim 8 , wherein the animal is inoculated with a vaccine containing the effective amount of the VLPs. 
     
     
         15 . The method of  claim 8 , wherein the immune response is effective to provide protection against viral challenge. 
     
     
         16 . The method of  claim 8 , wherein the effective amount of the VLPs is an amount effective to produce an immune response that protects the animal against challenge with live virus. 
     
     
         17 . A composition comprising purified VLPs produced by the method according to  claim 1 . 
     
     
         18 . The composition of  claim 17 , wherein the composition is a vaccine for comprising an effective amount of the VLPs. 
     
     
         19 . The method of  claim 18 , wherein the virus is selected from Swine Vesicular Disease Virus, Senecavirus A, Porcine Teschovirus, Bovine rhinitis A virus, Bovine rhinitis B virus, Equine rhinitis A virus, or Foot-and-mouth disease virus (FMDV) 
     
     
         20 . The composition of  claim 19 , further comprising an adjuvant.