Virus like particle purification
Methods for purifying human Calciviruses are disclosed, including Noroviruses and Sapoviruses.
1. A production method for Norovirus virus-like particles (VLPs), comprising:
transforming insect cells with a baculovirus vector containing a viral nucleic acid sequence encoding a Norovirus major capsid protein VP1;
cultivating the insect cells in a medium thereby expressing the VLPs in the medium;
stopping cultivation when the viability of the insect cells reaches 20% or less; and
isolating supernatant comprising the VLPs from the insect cells thus producing the VLPs.
2. The production method of claim 1 , wherein the Norovirus VLPs are Norovirus genogroup I VLPs.
3. The production method of claim 1 , wherein the Norovirus VLPs are Norovirus genogroup II VLPs.
4. The production method of claim 2 , wherein the Norovirus genogroup I VLPs are Norovirus genotype GI.1.
5. The production method of claim 3 , wherein the Norovirus genogroup II VLPs are Norovirus genotype GII.4.
6. The production method of claim 1 further comprising purifying the Norovirus VLPs.
7. The production method of claim 6 , wherein the purification of the Norovirus VLPs comprises using at least one chromatographic material.
8. The production method of claim 7 , wherein the at least one chromatographic material comprises hydroxyapatite chromatographic material.
9. The production method of claim 8 , wherein the Norovirus VLPs are eluted from the hydroxyapatite chromatographic material using a phosphate buffer.
10. The production method of claim 9 , wherein the phosphate buffer has a concentration of about 1 to about 300 mM.