Nucleic acid free ghost preparations
View Patent ↗The invention relates to preparations of bacterial ghosts which are substantially free of living bacterial cells and/or nucleic acids and their use in pharmaceutical preparations.
1. A preparation of bacterial ghosts which is substantially free of living bacterial cells, wherein the ratio of ghosts to living cells is at least 10 6 :1.
2. The preparation of claim 1 , wherein the ratio of ghosts to living cells is at least 10 8 :1.
3. A pharmaceutical composition comprising the preparation of claim 1 and a pharmaceutically acceptable carrier, diluent and/or adjuvant.
4. The composition of claim 3 which is a vaccine.
5. The composition of claim 3 which is an adjuvant.
6. The composition of claim 3 for use in human medicine.
7. The composition of claim 3 for use in veterinary medicine.
8. A method for obtaining a preparation of bacterial ghosts of claim 1 comprising the steps:
(a) providing bacterial cells comprising
(i) a gene encoding a lytic protein capable of forming a tunnel structure in the bacterial membranes and
(ii) a gene encoding an enzyme capable of hydrolyzing cytoplasmic components in the bacterial cells, wherein said enzyme is a nuclease,
(b) optionally cultivating the bacterial cells under conditions wherein the lytic gene and the enzyme gene are not expressed,
(c) subjecting the bacterial cells to conditions wherein the lytic gene and the enzyme gene are expressed and the cytoplasmic components of the bacterial cells are degraded and liberated and
(d) obtaining the resulting bacterial ghosts.
9. The method of claim 8 , wherein the gene encoding the lytic protein is the bacteriophage phiX174 gene E.
10. The method of claim 8 , wherein the nuclease is a Staphylococcus aureus nuclease.
11. The method of claim 8 , wherein the lytic gene and the enzyme gene are in operative linkage with a regulatable expression control sequence.
12. The method of claim 11 , wherein the lytic gene and the enzyme gene are each in operative linkage with a separate regulatable expression control sequence.
13. The method of claim 12 , wherein the expression of the lytic gene and the expression of the enzyme are induced at different times.
14. A preparation of bacterial ghosts which is substantially free of nucleic acids.
15. An isolated bacterial cell comprising
(i) a gene encoding a lytic protein capable of forming a tunnel structure in the bacterial membrane and
(ii) a gene encoding an enzyme capable of hydrolyzing cytoplasmic components in the bacterial cell,
wherein said lytic gene and/or enzyme gene are located on one or several plasmid vectors.