Invaplex from gram negative bacteria, method of purification and methods of use
AssigneeThe United States of America as represented by the Secretary of the Army
InventorsEdwin V. Oaks, Kevin Ross Turbyfill
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Abstract
Invaplex, a novel composition comprising invasin proteins and LPS from gram-negative bacteria is described as well as methods of using the novel composition as an adjuvant and a diagnostic tool.
Claims21
An immunogenic composition comprising an isolated lipopolysaccharide-protein complex (Invaplex) isolated from a water extract of gram-negative bacteria, wherein the complex is composed of at least one invasin protein associated with LPS of said gram-negative bacteria.
A method for preparing isolated Invaplex from Shigella, said method comprising the steps of:
- (i) extracting Shigella with water to form an aqueous phase having an immunogenic lipopolysaccharide-protein complex component, Invaplex,
- (ii) separating and discarding membrane fragments from said aqueous phase resulting in a solution containing the Invaplex;
- and
- (iii) isolating the Invaplex from said solution.
A method for preparing isolated Invaplex from Escherichia, said method comprising the steps of:
- (i) extracting Escherichia with water to form an aqueous phase having an immunogenic lipopolysaccharide-protein complex component, Invaplex;
- (ii) separating and discarding membrane fragments from said aqueous phase resulting in a solution containing the Invaplex;
- and
- (iii) isolating the Invaplex from the solution with an ion-exchange matrix.
A method for screening agents or drugs which reduce or eliminate Invaplex said method comprising detecting a dissociation of said Invaplex in the presence of said agent or drug.
A method for detecting gram-negative bacterial infection in a biological sample comprising
- (i) contacting a sample with a solid surface to which is attached an Invaplex isolated from bacteria suspected of causing the bacterial infection;
- and
- (ii) detecting the presence or absence of a complex formed between said isolated Invaplex and antibodies specific therefor in said sample wherein the presence of said complex indicates the presence of said bacterial infection, wherein the isolated Invaplex is an immunogenic lipopolysaccharide-protein complex that has been isolated from a water extract of the gram-negative bacteria, is an adjuvant and is composed of at least one invasin protein associated with LPS of the gram-negative bacteria.
A method to elicit an antigen-specific immune response in a subject, said method comprising administering to said subject an Invaplex isolated from a gram-negative bacteria along with said antigen wherein said antigen-specific immune response is chosen from the group consisting of cell-mediated immune response, humoral immune response, and mucosal immune response, wherein the isolated Invaplex is an immunogenic lipopolysaccharide-protein complex that is isolated from a water extract of gram-negative bacteria, is an adjuvant and is composed of at least one invasin protein associated with LPS of said gram-negative bacteria.
Dependencies are inferred from the claim text.