Methods for producing enhanced antigenic Helicobacter sp.
AssigneeAntex Biologics Inc.
InventorsJohn Lee Pace, Richard Ives Walker, Steven Michael Frey
Life of this patent
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Abstract
Methods using in vitro processes are disclosed for inducing or enhancing expression of enteric bacterial antigens or virulence factors. The methods, therefore, produce antigenically enhanced enteric bacteria. Also methods for using the antigenically enhanced bacteria are also disclosed, as well as vaccines containing the enteric bacteria. Specifically a whole enteric bacterium or components thereof are provided by Helicobacter species. Also there are other enteric bacteria which are useful for the disclosed invention; such as Campylobacter jejuni.
Claims19
A method of producing Helicobacter bacteria having enhanced antigenic properties which comprises growing a culture of the Helicobacter bacteria in vitro in a combination of conditions comprising:
- a) about 0.05% to about 3% bile or about 0.025% to about 0.6% of one or more bile acids or salts thereof;
- b) at a temperature between about 30.degree. C. and about 42.degree. C.;
- c) in air or an microaerophillic condition, wherein the microaerophillic condition comprises i) about 5% to about 20% CO.sub.2 with about 80% to about 95% air;
- or ii) about 5% to about 10% O.sub.2 with about 10% to about 20% CO.sub.2 with about 70% to about 85% N.sub.2 ;
- and
- d) a divalent cation chelator selected from the group consisting of 0 to about 100 .mu.M of BAPTA/AM, 0 to about 10 mM of EGTA, and 0 to about 100 .mu.M of EGTA/AM,
- for a sufficient time so that the culture is in a growth phase at about early log phase, between early log phase and stationary phase, or at about stationary phase.
A method of producing Helicobacter bacteria having enhanced antigenic properties which comprises growing a culture of the Helicobacter bacteria in vitro in a combination of conditions comprising:
- a) a divalent cation chelator selected from the group consisting of about 1.0 to about 25 .mu.M of BAPTA/AM, about 0.5 about 10 mM of EGTA, and about 1.0 to about 100 .mu.M of EGTA/AM;
- b) at a temperature between about 30.degree. C. and about 42.degree. C.;
- and
- c) in air or an microaerophillic condition, wherein the microaerophillic condition comprises i) about 5% to about 20% CO.sub.2 with about 80% to about 95% air;
- or ii) about 5% to about 10% O.sub.2 with about 10% to about 20% CO.sub.2 with about 70% to about 85% N.sub.2,
- for a sufficient time so that the culture is in a growth phase at about early log phase, between early log phase and stationary phase, or at about stationary phase.
A Helicobacter bacterium having enhanced antigenic properties, which is harvested from a culture of Helicobacter bacteria grown in vitro under a combination of conditions comprising:
- a) about 0.05% to about 3% bile or about 0.025% to about 0.6% of one or more bile acids or salts thereof;
- b) at a temperature between about 30.degree. C. and about 42.degree. C.;
- c) in air or an microaerophillic condition, wherein the microaerophillic condition comprises i) about 5% to about 20% CO.sub.2 with about 80% to about 95% air;
- or ii) about 5% to about 10% O.sub.2 with about 10% to about 20% CO.sub.2 with about 70% to about 85% N.sub.2 ;
- and
- d) a divalent cation chelator selected from the group consisting of 0 to about 25 .mu.M of BAPTA/AM, 0 to about 10 mM of EGTA, and 0 to about 100 .mu.M of EGTA/AM,
- wherein said Helicobacter culture is at about early log phase, between early log phase and stationary phase, or at about stationary phase.
A Helicobacter bacterium having enhanced antigenic properties, which is harvested from a culture of Helicobacter bacteria grown in vitro under a combination of conditions comprising:
- a) a divalent cation chelator, selected from the group consisting of about 1.0 to about 25 .mu.M of BAPTA/AM, about 0.5 to about 10 mM of EGTA, and about 1.0 to about 100 .mu.M of EGTA/AM;
- b) at a temperature between about 30.degree. and about 42.degree. C.;
- and
- c) in air or an microaerophillic condition, wherein the microaerophillic condition comprises i) about 5% to about 20% CO.sub.2 with about 80% to about 95% air;
- or ii) about 5% to about 10% O.sub.2 with about 10% to about 20% CO.sub.2 with about 70% to about 85% N.sub.2,
- wherein said Helicobacter culture is at about early log phase, between early log phase and stationary phase, or at about stationary phase.
Dependencies are inferred from the claim text.