NOVEL IMMUNOGENIC COMPOSITIONS FOR THE PREVENTION AND TREATMENT OF MENINGOCOCCAL DISEASE
The present invention relates to Neisseria ORF2086 proteins, crossreactive immunogenic proteins which can be isolated from nesserial strains or prepared recombinantly, including immunogenic portions thereof, biological equivalents thereof, antibodies that immunospecifically bind to the foregoing and nucleic acid sequences encoding each of the foregoing, as well as the use of same in immunogenic compositions that are effective against infection by Neisseria meningitidis serogroup B.
1 - 5 . (canceled)
6 . A composition comprising at least one isolated and purified protein having at least 95% amino acid sequence identity to a protein comprising the amino acid sequence of any of even numbered SEQ ID NOS:110-114.
7 . The composition of claim 6 , additionally comprising at least one protein comprising the amino acid sequence of any of even numbered SEQ ID NOS:176-252.
8 . (canceled)
9 . The composition of claim 6 , wherein said the at least one protein is nonpathogenic. free from any infectious impurities.
10 . The composition of claim 6 , wherein the at least one protein has a molecular weight of about 26,000 to about 30,000 daltons as measured by mass spectroscopy.
11 . The composition of claim 10 , wherein the at least one protein has a molecular weight of about 28-35 kDa as measure on a 10%-20% SDS polyacrylamide gel.
12 . The composition of claim 6 , wherein said composition additionally comprises a pharmaceutically acceptable buffer, diluent, adjuvant or carrier.
13 . The composition of claim 6 , wherein said composition additionally comprises a carrier.
14 . The composition of claim 6 , wherein said composition additionally comprises an adjuvant.
15 . The composition of claim 14 , wherein said adjuvant comprises a liquid.
16 . The composition of claim 6 , wherein the protein is a recombinant protein.
17 . The composition of claim 6 , wherein the protein is isolated from native Neisseria species.
18 . The composition of claim 6 , wherein the protein is a lipoprotein.
19 . The composition of claim 6 , wherein the protein is non-lipidated.
20 . The composition of claim 6 , wherein the composition additionally comprises at least one Neisseria species protein: PorA, PorB, transferrin binding protein, or opacity protein (Opc).
21 . The composition of claim 6 , wherein the composition additionally comprises at least one additional surface antigen of Neisseria species, said additional surface antigen being a non-ORF2086 protein.
22 . The composition of claim 6 , wherein said composition additionally comprises a polysaccharide.
23 . The composition of claim 6 , wherein said composition comprises an additional peptide, polypeptide or protein, said composition forming a conjugate that induces an immune response to two or more bacteria in a mammal.
24 - 59 . (canceled)
60 . A composition comprising:
(a) at least one isolated and purified protein comprising the amino acid sequence having at least 95% amino acid sequence identity to a protein comprising the amino acids of any of even numbered SEQ ID NOS:110-114; or
(b) at least one protein encoded by a polynucleotide that hydridizes under stingent conditions to a polynucleotide having at least 95% nucleic acid sequence identity to a polynucleotide comprising the nucleic acid sequence of any of odd numbered SEQ ID NOS:109-113.
61 . The composition of claim 60 , wherein the at least one protein comprises the amino acid sequence of any of even numbered SEQ ID NOS:110-114.
62 . The composition of claim 61 , additionally comprising at least one protein comprising the amino acid sequence of any of even numbered SEQ ID NOS:176-252.
63 - 71 . (canceled)
72 . The composition of claim 60 , wherein the composition additionally comprises at least one Neisseria species protein: Por A, Por B, transferrin binding protein, or opacity protein (Opc).
73 . The composition of claim 60 , wherein the composition additionally comprises at least one additional surface antigen of Neisseria species, said additional surface antigen being a non-ORF2086 protein.
74 . The composition of claim 60 , wherein the at least one protein has a molecular weight of about 26,000 to about 30,000 as measured by mass spectroscopy.
75 . The composition of claim 74 , wherein the at least one protein has a molecular weight of about 28-35 kDa as measured on a 10%-20% SDS polyacrylamide gel.
76 . The composition of claim 60 , wherein said composition additionally comprises a pharmaceutically acceptable buffer, diluent, adjuvant or carrier.
77 . The composition of claim 60 , wherein said composition additionally comprises a carrier.
78 . The composition of claim 60 , wherein said composition additionally comprises an adjuvant.
79 . The composition of claim 78 , wherein said adjuvant comprises a liquid.
80 . The composition of claim 60 , wherein the protein is non-lipidated.
81 . The composition of claim 60 , wherein the protein is a recombinant protein.
82 . The composition of claim 60 , wherein the protein is isolated from native Neisseria species.
83 . The composition of claim 60 , wherein the protein is a lipoprotein.
84 . The composition of claim 60 , wherein said composition additionally comprises a polysaccharide.
85 . The composition of claim 60 , wherein said composition comprises an additional peptide, polypeptide or protein, said composition forming a conjugate that induces an immune response to two or more bacteria in a mammal.
86 . A composition comprising: at least one isolated and purified protein from of a first bacterial strain of Neisseria species having at least 95% amino acid sequence identity to a protein comprising the amino acid sequence of any of even numbered SEQ ID NOS: 110-114; that provides immunogenicity against infection of a subject by a second bacterial strain of Neisseria species.
87 . The composition of claim 86 , wherein the first strain is a strain of Neisseria species and said second strain is a strain of Neisseria meningitidis serogroup B.
88 . The composition of claim 86 , wherein the first strain is any of strains L3 6275, CDC2369, CDC1034, L4 891, B16B6, W135 (ATCC35559), C11, Y(ATCC35561), M98 250732, M98 250771, CDC1135, M97 252153, CDC1610, CDC1492, L8 M978; M97 252988, M97 252697, 6557, 2996, M97 252976, M97 251854, CDC1521, M98 250622, 870446, M97 253248, M98 250809, L5 M981, NMB or M98 250572.
89 . (canceled)
90 . The composition of claim 86 , wherein the protein is a recombinant protein.
91 . The composition of claim 86 , wherein the protein is isolated from native Neisseria species.
92 . The composition of claim 86 , wherein the protein is a lipoprotein.
93 . The composition of claim 86 , wherein said composition additionally comprises a pharmaceutically acceptable buffer, diluent, adjuvant or carrier.
94 . The composition of claim 86 , wherein said composition additionally comprises a carrier.
95 . The composition of claim 86 , wherein said composition additionally comprises an adjuvant.
96 . The composition of claim 95 , wherein said adjuvant comprises a liquid.
97 . The composition of claim 86 , wherein the protein is non-lipidated.
98 . The composition of claim 86 , wherein said composition additionally comprises a polysaccharide.
99 . The composition of claim 86 , wherein said composition comprises an additional peptide, polypeptide or protein, said composition forming a conjugate that induces an immune response to two or more pathogens in a mammal.
100 - 156 . (canceled)
157 . A composition comprising:
(a) at least one polynucleotide that encodes at least one of:
(i) at least one isolated and purified protein having at least 95% amino acid sequence identity to a protein comprising the amino acid sequence of any of even numbered SEQ ID NOS:110-114; or
(ii) at least one isolated and purified protein encoded by a polynucleotide that hydridizes under stringent conditions to a polynucleotide having at least 95% nucleic acid sequence identity to a polynucleotide comprising the nucleic acid sequence of any of odd numbered SEQ ID NOS:109-113.
(b) at least one polynucleotide that hybridizes under stringent conditions to any of the polynucleotides described in (a).
158 . The composition of claim 157 , wherein said at least one polynucleotide comprises the nucleic acid sequence of any of the odd numbered SEQ ID NOS: 109-113.
159 . The composition of claim 157 , wherein said composition comprises a vector.
160 . The composition of claim 157 , wherein the stringent conditions are high stringency southern hybridization conditions.
161 . The composition of claim 157 , wherein said composition additionally comprises a nucleic acid sequence encoding for an additional peptide, polypeptide or protein.
162 . A composition comprising:
(a) at least one polynucleotide that encodes at least one protein of a first bacterial strain of Neisseria species that provides immunogenicity against infection of a subject by a second bacterial strain of Neisseria species; or
(b) at least one polynucleotide that hybridizes under stringent conditions to at least one polynucleotide of (a).
163 . The composition of claim 162 , wherein said at least one isolated polynucleotide comprises the nucleic acid sequence of any of the odd numbered SEQ ID NOS: 109-113.
164 . The composition of claim 162 , additionally comprising a P4 leader sequence (SEQ ID NO. 322).
165 . The composition of claim 162 , wherein said composition comprises a vector.
166 . The composition of claim 162 , wherein the stringent conditions are high stringency southern hybridization conditions.
167 . The composition of claim 162 , wherein said composition additionally comprises a nucleic acid sequence encoding for an additional peptide, polypeptide or protein.
168 - 175 . (canceled)
176 . A composition comprising:
a vector comprising any of:
(a) at least one polynucleotide that encodes at least one of the polypeptides having at least 95% amino acid sequence identity to a protein comprising the amino acids sequence of any of the even numbered SEQ ID NOS: 110-114; or
(b) at least one polynucleotide that hybridizes under stringent conditions to at least one polynucleotide of (a).
177 . The composition of claim 176 , wherein the vector comprises the nucleic acid sequence of any of odd numbered SEQ ID NOS:109-113.
177 . The composition of claim 176 , wherein the vector is a plasmid.
178 . The composition of claim 176 , wherein the vector is a phage.
179 . The composition of claim 176 , wherein the vector is a bacteriophage.
180 . The composition of claim 176 , wherein the vector is a moderate phage.
181 - 182 . (canceled)
183 . A composition prepared by a process comprising:
expressing in a host cell a nucleic acid sequence encoding:
at least one isolated and purified protein encoded by an open reading frame, ORF 2086 of Neisseria species, said open reading frame encoding the at least one protein comprising an amino acid sequence of any of even numbered SEQ ID NOs: 110-114 that provides immunogenicity against infection by Neisseria meningitidis serogroup B in a subject.
184 . The composition of claim 183 , wherein the nucleic acid sequence is any of the odd numbered SEQ ID NOS: 109-113.
185 . The composition of claim 183 , wherein the nucleic acid sequence encodes a protein comprising any of the even numbered SEQ ID NOS: 110-114.
186 . (canceled)
187 . The composition of claim 183 , wherein the protein has a molecular weight of about 26,000 to about 30,000 as measured by mass spectroscopy.
188 . The composition of claim 187 , wherein the protein has a molecular weight of about 28-35 kDa as measured on a 10%-20% SDS polyacrylamide gel.
189 . The composition of claim 183 , wherein said composition additionally comprises a pharmaceutically acceptable buffer, diluent, adjuvant or carrier.
190 . The composition of claim 183 , wherein said composition additionally comprises a carrier.
191 . The composition of claim 183 , wherein said composition additionally comprises an adjuvant.
192 . The composition of claim 191 , wherein the adjuvant is a liquid.
193 . The composition of claim 183 , wherein the protein is a lipidated protein.
194 . (canceled)
195 . A composition prepared by a process comprising: isolating and purifying from Neisseria species at least one isolated and purified protein encoded by an open reading frame, ORF 2086 of the Neisseria species, said open reading frame encoding the at least one protein encoded by a polynucleotide sequence having at least 95% identity to the polynucleotide that comprises the nucleic acid sequence of any of the odd numbered SEQ ID NOS: 109-113.
196 . The composition of claim 195 , wherein the process further comprises introducing a non-native leader sequence to the at least one isolated polynucleotide.
197 . The composition of claim 196 , wherein the non-native leader sequence is P4 leader sequence (SEQ ID NO. 322).
198 - 215 . (canceled)
216 . A composition prepared by a process comprising expressing in a host cell a nucleic acid sequence having at least 95% nucleic acid sequence identity to a polynucleotide comprising a nucleic acid sequence of any of the odd numbered SEQ ID NOS: 109-113.
217 . The composition of claim 216 wherein the nucleic acid sequence encodes a protein having at least 95% amino acid sequence identity to a protein comprising an amino acid sequence of comprising any of the even numbered SEQ ID NOS: 110-114.
218 . The composition of claim 216 wherein the protein has a molecular weight of about 26,000 to about 30,000 as measured by mass spectroscopy.
219 . The composition of claim 218 , wherein the protein has a molecular weight of about 28-35 kDa as measured on a 10%-20% SDS polyacrylamide gel.
220 . The composition of claim 216 , wherein said composition additionally comprises a pharmaceutically acceptable buffer, diluent, adjuvant or carrier.
221 . The composition of claim 216 wherein said composition additionally comprises a carrier.
222 . The composition of claim 216 , wherein said composition additionally comprises an adjuvant.
223 . The composition of claim 222 , wherein said adjuvant comprises a liquid.
224 . The composition of claim 216 wherein nucleic the stringent conditions are high stringency southern hybridization conditions.
225 . A composition prepared by a process comprising:
isolating and purifying from Neisseria species any of:
(a) at least one polynucleotide that encodes at least one of the polypeptides having at least 95% amino acid sequence identity to a protein comprising the amino acid sequence of any of the even numbered SEQ ID NOS: 110-114; or
(b) at least one polynucleotide that hybridizes under stringent conditions to at least one polynucleotide of (a).
226 . The composition of claim 225 , wherein the at least one polynucleotide comprises the nucleic acid sequence of any of the odd numbered SEQ ID NOS: 109-113.
227 . The composition of claim 225 , wherein the process further comprises introducing a non-native leader sequence to the at least one polynucleotide.
228 . The composition of claim 227 , wherein the non-native leader sequence is P4 leader sequence (SEQ ID NO. 322).
229 . A composition prepared by a process comprising:
isolating and purifying from Neisseria species any of:
(a) at least one protein having at least 95% amino acid sequence identity to a protein comprising the amino acid sequence of any of even numbered SEQ ID NOS:110-114;or
(b) at least one protein encoded by a polynucleotide that hydridizes under stingent conditions to a polynucleotide having at least 95% nucleic acid sequence identity to a polynucleotide comprising a nucleic acid sequence of any of odd numbered SEQ ID NOS:109-113.
230 . The composition of claim 229 , wherein the protein has a molecular weight of about 26,000 to about 30,000 as measured by mass spectroscopy.
231 . The composition of claim 230 , wherein the protein has a molecular weight of about 28-35 kDa as measured on a 10%-20% SDS polyacrylamide gel.
232 . The composition of claim 229 , wherein said composition additionally comprises a pharmaceutically acceptable buffer, diluent, adjuvant or carrier.
233 . The composition of claim 229 , wherein said composition additionally comprises a carrier.
234 . The composition of claim 229 , wherein said composition additionally comprises an adjuvant.
235 . The composition of claim 234 , wherein said adjuvant comprises a liquid.
236 . The composition of claim 229 , wherein the stringent conditions are high stringency southern hybridization conditions.
237 . A composition prepared by a process comprising:
inserting into a vector any of:
(a) at least one isolated polynucleotide that encodes at least one polypeptide having at least 95% amino acid sequence identity to a protein comprising the amino acid sequence of any of the even numbered SEQ ID NOS: 110-114; or
(b) at least one isolated polynucleotide that hybridizes under stringent conditions to at least one polynucleotide of (a).
238 . The composition of claim 237 , wherein the vector is a plasmid.
239 . The composition of claim 237 , wherein the vector is a phage.
240 . The composition of claim 237 , wherein the vector is a bacteriophage.
241 . The composition of claim 237 , wherein the vector is a moderate phage.
242 . The composition of claim 237 , further comprising a non-native leader sequence
243 . The composition of claim 242 , wherein the non-native leader sequence is P4 leader sequence (SEQ ID NO. 322).
244 - 315 . (canceled)
316 . A method of preparing a composition comprising:
expressing in a host cell a nucleic acid sequence encoding any of:
(a) at least one protein having at least 95% amino acid sequence identity to a protein comprising the amino acid sequence any of even numbered SEQ ID NOS:110-114; or
(b) at least one protein encoded by a polynucleotide that that hydridizes under stingent conditions to a polynucleotide having at least 95% nucleic acid identity to a polynucleotide comprising a nucleic acid sequence any of odd numbered SEQ ID NOS:109-113.
317 . The method of claim 316 , wherein the nucleic acid sequence is expressed in vivo.
318 . The method of claim 316 , wherein the nucleic acid sequence is expressed in vitro.
319 . The method of claim 316 , wherein the vector is a plasmid.
320 . The method of claim 316 , wherein the vector is a phage.
321 . The method of claim 316 , further comprising associating a non-native leader sequence with said at least one isolated polynucleotide.
322 . The method of claim 316 , wherein the non-native leader sequence is P4 leader sequence (SEQ ID NO. 267).
323 - 325 . (canceled)
326 . A method of preparing a composition comprising:
isolating and purifying from Neisseria species any of:
(a) at least one protein having at least 95% amino acid sequence identity to a protein comprising the amino acid sequence of any of the even numbered SEQ ID NOS: 110-114or
(b) at least one protein encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide having at least 95% nucleic acid sequence identity to a polynucleotide comprising the nucleic acid sequence of any of the odd numbered SEQ ID NOS: 109-113.
327 . The method of claim 326 , wherein the stringent conditions are high stringency southern hybridization conditions.
328 - 342 . (canceled)
343 . An isolated and purified polynucleotide comprising:
at least one polynucleotide that encodes:
(a) at least one isolated and purified protein having at least 95% amino acid sequence identity to a protein comprising the amino acid sequence of any of even numbered SEQ ID NOS: 110-114; or
(b) at least one isolated and purified protein encoded by a polynucleotide that hydridizes under stringent conditions to a polynucleotide having at least 95% nucleic acid sequence identity to a polynucleotide comprising the nucleic acid sequence of any of odd numbered SEQ ID NOS: 109-113.
344 . An isolated and purified polypeptide comprising:
(a) an amino acid sequence comprising at least one isolated and purified protein having at least 95% amino acid sequence identity to a protein comprising the amino acid sequence of any of even numbered SEQ ID NOS: 110-114;or
(b) an amino acid sequence that is encoded by a polynucleotide having at least 95% nucleic acid sequence identity to a polynucleotide comprising the nucleic acid sequence of any of the odd numbered sequences of SEQ ID NOS 109-113.
345 . An isolated an purified polynucleotide comprising the nucleic acid sequence of any of odd numbered SEQ ID NOS 109-113.