Methods and nucleic acid molecules for aav vector selection
US-2024417717-A1 · Dec 19, 2024 · US
US9109014B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9109014-B2 |
| Application number | US-201013503828-A |
| Country | US |
| Kind code | B2 |
| Filing date | Nov 2, 2010 |
| Priority date | Nov 2, 2009 |
| Publication date | Aug 18, 2015 |
| Grant date | Aug 18, 2015 |
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Provided herein is a nucleic acid comprising consensus amino acid sequence of foot-and-mouth disease FMDV VP1-4 coat proteins of FMDV subtypes A, Asia 1, C, O, SAT1, SAT2, and SAT3 as well as plasmids and vaccines expressing the sequences. Also provided herein is methods for generating an immune response against one or more FMDV subtypes using the vaccine as described above as well as methods for deciphering between vaccinated mammals with the vaccine and those that are infected with FMDV.
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The invention claimed is: 1. A pharmaceutical composition comprising a nucleic acid molecule comprising a nucleic acid sequence that encodes a protein selected from the group consisting of: a protein comprising SEQ ID NO: 40, with or without a leader sequence; and a protein with 95% identity to SEQ ID NO: 40. 2. The pharmaceutical composition of claim 1 , wherein the nucleic acid molecule comprises a nucleic acid sequence selected from the group consisting of: SEQ ID NO: 39 with or without coding sequence for a leader sequence; and nucleic acid sequences with 95% or more identity to SEQ ID NO: 39. 3. The pharmaceutical composition of claim 1 comprising a nucleic acid sequence that encodes SEQ ID NO: 40 with or without a leader sequence. 4. The pharmaceutical composition of claim 3 comprising SEQ ID NO: 39. 5. The pharmaceutical composition of claim 1 wherein the leader sequence is an IgE leader sequence. 6. The pharmaceutical composition of claim 1 wherein the nucleic acid is a plasmid. 7. The pharmaceutical composition of claim 1 wherein said nucleic acid is a plasmid that is an expression vector. 8. A method of eliciting an immune response against one or more FMDV virus subtypes in a mammal, comprising administering a pharmaceutical composition according to claim 1 . 9. The method of claim 8 , wherein the nucleic acid molecule is a DNA plasmid, the method comprising the steps of: a) administering the nucleic acid molecule to the tissue of the mammal; and b) electroporating cells of the tissue with a pulse of energy at a constant current effective to permit entry of the DNA plasmid into the cells. 10. The method of claim 9 , wherein step a) comprises injecting the DNA plasmid vaccine into intradermic, subcutaneous, or muscle tissue. 11. The method of claim 9 , wherein the current is preset for delivering to the tissue and pulse of energy is at a constant current that equals the preset current. 12. The method of claim 9 , wherein the electroporating step further comprises: a) measuring the impedance in the electroporated cells; b) adjusting the energy level of the pulse of energy relative to the measured impedance to maintain a constant current in the electroporated cells; wherein the measuring and adjusting steps occur within a lifetime of the pulse of energy. 13. The method of claim 9 , wherein the electroporation step comprise delivering the pulse of energy to a plurality of electrodes according to a pulse sequence pattern that delivers the pulse of energy in a decentralized pattern. 14. The pharmaceutical composition of claim 1 further comprising one or more additional nucleic acid sequences selected from the group consisting of: nucleic acid sequences that encode a protein selected from the group of SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38 and 42 with or without a leader sequence; and nucleic acid sequences that encode proteins selected from the group consisting of proteins that have 95% or more identity to a protein selected from the group consisting of SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38 and 42. 15. The pharmaceutical composition of claim 14 , wherein the one or more additional nucleic acid sequences are selected from the group consisting of: nucleic acid sequences that encode a protein selected from the group of SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38 and 42 with or without a leader sequence. 16. The pharmaceutical composition of claim 14 , wherein the one or more additional nucleic acid sequences are selected from the group consisting of: SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 and 41, with or without coding sequence for a leader sequence, and nucleic acid sequences that are 95% or more identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 and 41. 17. The pharmaceutical composition of claim 16 , wherein the one or more additional nucleic acid sequences are selected from the group consisting of: SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 and 41, with or without coding sequence for a leader sequence. 18. The pharmaceutical composition of claim 17 , wherein said composition comprises a nucleic acid sequence that encodes SEQ ID NO: 40 and SEQ ID NO: 42. 19. The pharmaceutical composition of claim 18 , wherein said composition comprises a nucleic acid sequence that comprises SEQ ID NO: 39 and SEQ ID NO: 41. 20. A pharmaceutical composition comprising a molecule comprising a nucleic acid sequence that encodes a protein comprising SEQ ID NO: 40, with or without a leader sequence. 21. The pharmaceutical composition of claim 20 , wherein the nucleic acid sequence comprises SEQ ID NO: 39 with or without coding sequence for a leader sequence.
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