Rabies glycoprotein virus-like particles (VLPS)
The present invention is generally related to virus-like particles (VLPs) comprising rabies virus (RV) glycoproteins (G proteins) and methods for making and using them, including immunogenic compositions such as vaccines for the treatment and/or prevention of rabies virus infection.
1. A method of making a micelle particle comprising rabies virus (RV) glycoprotein (G protein), wherein the G proteins form a trimer, comprising:
(a) transforming an Sf9 cell to express a nucleic acid encoding a RV G protein; and
(b) culturing said Sf9 cell under conditions conducive to the production of said RV G protein trimers; and
(c) purifying a micelle particle comprising the RV G protein trimers in the presence of NP-9 detergent.
2. The method according to claim 1 , wherein said Sf9 cell is transfected with a baculovirus vector comprising a nucleic acid comprising SEQ ID NO:1.
3. The method according to claim 1 , wherein said Sf9 cell is transfected with a baculovirus vector comprising a nucleic acid that encodes a RV G protein comprising SEQ ID NO:2.
4. The method according to claim 1 , wherein step (c) comprises lentil lectin affinity chromatography.
5. The method according to 1 , wherein step (c) comprises:
(a) anion exchange chromatography;
(b) cation exchange chromatography; and
(c) lentil lectin affinity chromatography.
6. The method according to claim 5 , wherein the anion exchange chromatography utilizes an elution buffer comprising 25 mM Tris pH 8.0, 180 mM NaCl, and 0.02% NP-9.
7. The method according to claim 5 , wherein the cation exchange chromatography utilizes an elution buffer comprising 25 mM NaHPO 4 pH 6.8, 300 mM NaCl, and 0.02% NP-9.
8. The method according to claim 5 , wherein the lentil lectin affinity chromatography utilizes an elution buffer comprising 25 mM NaHPO 4 pH 6.8, 50 mM NaCl, 500 mM Methyl-α-D-mannopyranoside and 0.02% NP-9.
9. The method according to claim 1 , wherein the G protein trimer forms spikes on the surface of the micelle particle.
10. The method according to claim 1 , wherein the RV G protein is derived from an RV strain selected from human, canine, bat, raccoon, skunk, and fox.
11. The method according to claim 1 , wherein the micelle particle is stored in a buffer comprising 25 mM NaHPO 4 pH 6.8, 300 mM NaCl, and 0.02% NP-9.
12. A pharmaceutical composition comprising the micelle particle comprising rabies virus (RV) glycoprotein (G protein), wherein the G proteins form a trimer; 25 mM NaHPO 4 pH 6.8, 300 mM NaCl, and 0.02% NP-9.
13. The composition of claim 12 further comprising an adjuvant.
14. The composition of claim 13 , wherein the adjuvant is alum.