IP Library › Granted Patent US 9,981,026

Granted Patent B2

US 9,981,026 · App. 15/370,797 · Granted May 29, 2018

Compositions produced using enteric pathogens and methods of use

Inventors: Daryll A. Emery (New London, MN); Darren E. Straub (New London, MN); Laura Wonderling (Des Moines, IA)

Assignee: EPITOPIX, LLC

A61K39/0258 A61K39/105 C07K14/205 C07K14/245 A61K2039/54 A61K2039/542 A61K2039/545 A61K2039/55 A61K2039/552 Y10S530/825

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Quick Facts

Patent No.: US 9,981,026
App. No.: 15/370,797
Granted: May 29, 2018
Kind: B2

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Abstract

The present invention provides compositions including polypeptides having the characteristics of polypeptides expressed by a reference microbe such E. coli or Salmonella . Examples of Salmonella strains that can be used include, for instance, S. enterica serovar Newport, S. enterica serovar Enteritidis, S. enterica serovar Typhimurium, and S. enterica serovar Dublin. Also provided are compositions including polypeptides having a particular molecular weight and a mass fingerprint that includes polypeptide fragments having a particular set of masses, or polypeptides having an amino acid sequence with at least about 95% identity with an amino acid sequence, wherein the polypeptide has seroreactive activity. The present invention also provides methods of making and methods of using such compositions.

Claims (53)

1. A composition comprising:

a first isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 90 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the first isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having m/z values of 629.7±300 parts per million (ppm), 772.6±300 ppm, 831.5±300 ppm, 991.6±300 ppm, 1178.6±300 ppm, 1285.6±300 ppm, 1321.7±300 ppm, 1369.7±300 ppm, 1433.8±300 ppm, 1516.8±300 ppm, 1619.9±300 ppm, 1634.9±300 ppm, 1706.9±300 ppm, 1788.0±300 ppm, 1798.0±300 ppm, 1872.1±300 ppm, 1966.2±300 ppm, 1982.1±300 ppm, 2089.1±300 ppm, 2175.1±300 ppm, 2303.2±300 ppm, 2601.5±300 ppm, 2706.6±300 ppm, 2844.6±300 ppm, 3082.8±300 ppm, and 3213.8±300 ppm;

a second isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 86 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the second isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having m/z values of 976.5±300 ppm, 992.6±300 ppm, 1095.5±300 ppm, 1247.6±300 ppm, 1278.7±300 ppm, 1359.7±300 ppm, 1396.5±300 ppm, 1436.7±300 ppm, 1493.6±300 ppm, 1572.9±300 ppm, 1650.8±300 ppm, 1665.8±300 ppm, 1810.9±300 ppm, 1914.0±300 ppm, 2134.3±300 ppm, 2211.2±300 ppm, and 2270.1±300 ppm;

a third isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 83 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the third isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having m/z values of 837.27±300 ppm, 884.29±300 ppm, 1048.20±300 ppm, 1127.26±300 ppm, 1338.26±300 ppm, 1351.29±300 ppm, 1397.26±300 ppm, 1472.33±300 ppm, 1621.46±300 ppm, 1650.47±300 ppm, 1722.49±300 ppm, 1727.41±300 ppm, 1759.47±300 ppm, 1813.46±300 ppm, 1829.43±300 ppm, 2282.75±300 ppm, and 2512.79±300 ppm;

a fourth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 79 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the fourth isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having m/z values of 716.5±300 ppm, 952.5±300 ppm, 1135.5±300 ppm, 1155.7±300 ppm, 1336.6±300 ppm, 1396.6±300 ppm, 1447.7±300 ppm, 1512.7±300 ppm, 1532.7±300 ppm, 1653.8±300 ppm, 1657.8±300 ppm, 1667.9±300 ppm, 1717.9±300 ppm, 1780.0±300 ppm, 1861.0±300 ppm, 1964.1±300 ppm, 2262.2±300 ppm, and 2398.2 Da±300 ppm;

a fifth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 66 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the fifth isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having m/z values of 632.33±450 ppm, 945.20±450 ppm, 1191.31±450 ppm, 1238.16±450 ppm, 1440.25±450 ppm, 1561.26±450 ppm, 1647.24±450 ppm, 1792.36±450 ppm, 2062.63±450 ppm, 2085.55±450 ppm, 2190.41±450 ppm, 2248.51±450 ppm, 2454.66±450 ppm, and 2628.63±450 ppm;

a sixth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 66 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the sixth isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having m/z values of 679.45±450 ppm, 1295.14±450 ppm, 1299.04±450 ppm, 1304.17±450 ppm, 1423.20±450 ppm, 1550.34±450 ppm, 1820.33±450 ppm, 1892.48±450 ppm, 1918.33±450 ppm, 2159.57±450 ppm, 2323.52±450 ppm, 2357.55±450 ppm, and 2699.58±450 ppm; and

an effective amount of a pharmaceutically acceptable adjuvant.

2. The composition of claim 1 further comprising

a seventh isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 56 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the seventh isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having m/z values of 1285.15±450 ppm, 1395.19±450 ppm, 1550.25±450 ppm, 1616.35±450 ppm, 1829.34±450 ppm, 2034.42±450 ppm, 2139.53±450 ppm, 2183.52±450 ppm, and 2242.47±450 ppm;

an eighth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 38 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the eighth isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having m/z values of 705.5±450 ppm, 885.4±450 ppm, 931.5±450 ppm, 939.4±450 ppm, 1120.4±450 ppm, 1122.5±450 ppm, 1124.4±450 ppm, 1171.4±450 ppm, 1290.4±450 ppm, 1348.5±450 ppm, 1379.4±450 ppm, 1380.5±450 ppm, 1439.5±450 ppm, 1664.6±450 ppm, 1820.7±450 ppm, 1821.7±450 ppm, 2232.9±450 ppm, 2353.9±450 ppm, 2447.9±450 ppm, 2585.1±450 ppm, 2792.1±450 ppm, 2991.3±450 ppm, and 3106.3±450 ppm;

a ninth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 37 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the ninth isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having m/z values of 818.4±300 ppm, 915.4±300 ppm, 1207.5±300 ppm, 1055.3±300 ppm, 1083.4±300 ppm, 1155.4±300 ppm, 1222.5±300 ppm, 1280.4±300 ppm, 1378.5±300 ppm, 1409.4±300 ppm, 1443.5±300 ppm, 1565.5±300 ppm, 1654.6±300 ppm, 1709.7±300 ppm, 2231.9±300 ppm, 2600.1±300 ppm, 2601.0±300 ppm, 2671.0±300 ppm, and 3478.4±300 ppm; or

a tenth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 29 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the tenth isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having m/z values of 951.32±300 ppm, 1020.31±300 ppm, 1485.37±300 ppm, 1501.36±300 ppm, 1517.50±300 ppm, 1676.54±300 ppm, and 1692.47±300 ppm.

3. A method of inducing production in an animal of antibody that specifically binds an iron regulated polypeptide isolated from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, the method comprising:

administering to the animal an effective amount of the composition of claim 1 .

4. The method of claim 3 wherein the animal is avian, bovine, caprine, ovine, porcine, bisontine, cervine , equine, a companion animal, or human.

5. The method of claim 4 wherein the method further comprises collecting the antibody from the animal.

6. The method of claim 5 wherein the method further comprises isolating the antibody.

7. A composition comprising:

an effective amount of a pharmaceutically acceptable adjuvant.

8. The composition of claim 7 further comprising:

a seventh isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 56 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the seventh isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having the amino acid sequences of SEQ ID NO:822 to SEQ ID NO:830;

a tenth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 29 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the tenth isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having the amino acid sequences of SEQ ID NO:873 to SEQ ID NO:875, SEQ ID NO:877, and SEQ ID NO:878.

9. A method of inducing production in an animal of antibody that specifically binds an iron regulated polypeptide isolated from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, the method comprising:

administering to the animal an effective amount of the composition of claim 7 .

10. The method of claim 9 wherein the animal is avian, bovine, caprine, ovine, porcine, bisontine, cervine , equine, a companion animal, or human.

11. The method of claim 6 wherein the method further comprises collecting the antibody from the animal.

12. The method of claim 11 wherein the method further comprises isolating the antibody.

13. A composition comprising:

a first isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 90 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the first isolated iron-regulated polypeptide comprises the amino acid sequences of SEQ ID NO:716, SEQ ID NO:717, SEQ ID NO:718, SEQ ID NO:719, SEQ ID NO:720, SEQ ID NO:721, SEQ ID NO:722, SEQ ID NO:724, SEQ ID NO:725, SEQ ID NO:726, SEQ ID NO:727, SEQ ID NO:728, SEQ ID NO:729, SEQ ID NO:730, SEQ ID NO:731, SEQ ID NO:732, SEQ ID NO:733, SEQ ID NO:734, SEQ ID NO:735, SEQ ID NO:738, SEQ ID NO:739, SEQ ID NO:740, and SEQ ID NO:741,

a second isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 86 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the second isolated iron-regulated polypeptide comprises the amino acid sequences of SEQ ID NO:742, SEQ ID NO:743, SEQ ID NO:744, SEQ ID NO:745, SEQ ID NO:746, SEQ ID NO:747, SEQ ID NO:748, SEQ ID NO:749, SEQ ID NO:750, SEQ ID NO:751, SEQ ID NO:753, SEQ ID NO:754, SEQ ID NO:755, SEQ ID NO:756, SEQ ID NO:757, and SEQ ID NO:758;

a third isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 83 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the third isolated iron-regulated polypeptide comprises the amino acid sequences of SEQ ID NO:759, SEQ ID NO:760, SEQ ID NO:761, SEQ ID NO:763, SEQ ID NO:764, SEQ ID NO:765, SEQ ID NO:766, SEQ ID NO:767, SEQ ID NO:768, SEQ ID NO:769, SEQ ID NO:770, SEQ ID NO:771, SEQ ID NO:772, SEQ ID NO:774, and SEQ ID NO:775;

a fourth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 79 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the fourth isolated iron-regulated polypeptide comprises the amino acid sequences of SEQ ID NO:776, SEQ ID NO:777, SEQ ID NO:778, SEQ ID NO:779, SEQ ID NO:780, SEQ ID NO:781, SEQ ID NO:782, SEQ ID NO:783, SEQ ID NO:784, SEQ ID NO:785, SEQ ID NO:786, SEQ ID NO:787, SEQ ID NO:788, SEQ ID NO:789, SEQ ID NO:790, SEQ ID NO:791, SEQ ID NO:792, SEQ ID NO:793, and SEQ ID NO:794;

a fifth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 66 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the fifth isolated iron-regulated polypeptide comprises the amino acid sequences of SEQ ID NO:795, SEQ ID NO:796, SEQ ID NO:797, SEQ ID NO:798, SEQ ID NO:799, SEQ ID NO:800, SEQ ID NO:801, SEQ ID NO:802, SEQ ID NO:803, SEQ ID NO:804, SEQ ID NO:805, SEQ ID NO:806, SEQ ID NO:807, and SEQ ID NO:808;

a sixth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 66 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the sixth isolated iron-regulated polypeptide comprises the amino acid sequences of SEQ ID NO:809, SEQ ID NO:810, SEQ ID NO:811, SEQ ID NO:812, SEQ ID NO:814, SEQ ID NO:815, SEQ ID NO:816, SEQ ID NO:817, SEQ ID NO:818, SEQ ID NO:819, SEQ ID NO:820, and SEQ ID NO:821; and

an effective amount of a pharmaceutically acceptable adjuvant.

14. The composition of claim 13 further comprising:

a seventh isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 56 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the seventh isolated iron-regulated polypeptide comprises the amino acid sequences of SEQ ID NO:822, SEQ ID NO:823, SEQ ID NO:824, SEQ ID NO:825, SEQ ID NO:826, SEQ ID NO:827, SEQ ID NO:828, SEQ ID NO:829, and SEQ ID NO:830;

an eighth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 38 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the eighth isolated iron-regulated polypeptide comprises the amino acid sequences of SEQ ID NO:831, SEQ ID NO:832, SEQ ID NO:833, SEQ ID NO:834, SEQ ID NO:835, SEQ ID NO:836, SEQ ID NO:837, SEQ ID NO:838, SEQ ID NO:839, SEQ ID NO:840, SEQ ID NO:841, SEQ ID NO:843, SEQ ID NO:844, SEQ ID NO:847, SEQ ID NO:848, SEQ ID NO:851, SEQ ID NO:853, and SEQ ID NO:852;

a ninth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 37 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the ninth isolated iron-regulated polypeptide comprises the amino acid sequences of SEQ ID NO:854, SEQ ID NO:855, SEQ ID NO:856, SEQ ID NO:857, SEQ ID NO:858, SEQ ID NO:859, SEQ ID NO:860, SEQ ID NO:861, SEQ ID NO:863, SEQ ID NO:864, SEQ ID NO:866, SEQ ID NO:868, SEQ ID NO:869, SEQ ID NO:870, and SEQ ID NO:872; or

a tenth isolated iron-regulated polypeptide from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, having a molecular weight of 29 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the tenth isolated iron-regulated polypeptide comprises the amino acid sequences of SEQ ID NO:873, SEQ ID NO:874, SEQ ID NO:875, SEQ ID NO:877, and SEQ ID NO:878.

15. A method of inducing production in an animal of antibody that specifically binds an iron regulated polypeptide isolated from E. coli strain BEcO157(stx-), deposited under ATCC accession number PTA-124618, the method comprising administering to the animal an effective amount of the composition of claim 13 .

16. The method of claim 15 wherein the animal is avian, bovine, caprine, ovine, porcine, bisontine, cervine , equine, a companion animal, or human.

17. The method of claim 16 wherein the method further comprises collecting the antibody from the animal.

18. The method of claim 17 wherein the method further comprises isolating the antibody.

Assignments (6)

ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 2, 2024

From: EPITOPIX, LLC

To: VAXXINOVA US, INC.

Reel/Frame 066488/0734 →

ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 21, 2016

From: EMERY, DARYLL A.; STRAUB, DARREN E.

To: EPITOPIX, LLC

Reel/Frame 041090/0681 →