Claims for Patent: 10,011,843

Summary for Patent: 10,011,843

Title:	Biological indicator
Abstract:	This invention relates to a biological indicator derived from a composition comprising: a host organism comprising a spore forming bacteria; a reporter gene for producing an indicator enzyme; a regulatory gene; and a vehicle for inserting the reporter gene and the regulatory gene in the host organism; the host organism bearing a transposable genetic element in its genome for inserting an insertion sequence in the regulatory gene; the insertion sequence comprising a transposase, a pair of terminal inverted repeat sequences, and at least one open reading frame for expressing the transposase. The vehicle may be taken up by the host organism. The insertion sequence may be inserted in the regulatory gene. The host organism may undergo sporulation to form the biological indicator. A process and an apparatus for using the biological indicator are disclosed.
Inventor(s):	Franciskovich; Phillip P. (Concord, OH), Cregger; Tricia A. (Fairlawn, OH), Yirava; William A. (Willoughby, OH), Burke; Peter A. (Concord, OH)
Assignee:	American Sterilizer Company (Mentor, OH)
Application Number:	15/618,156
Patent Claims:	1. A composition comprising: a host organism comprising a spore forming bacteria selected from the group consisting of Geobacillus stearothermophilus, Bacillus atrophaeus, Bacillus sphaericus, Bacillus anthracis, Bacillus pumilus, Bacillus coagulans, Bacillus subtilis globigii, Bacillus cereus, or Bacillus circulans; a reporter gene for producing an indicator enzyme; a regulatory gene; and a vehicle for inserting the reporter gene and the regulatory gene in the host organism; the host organism bearing a transposable genetic element in its genome for inserting an insertion sequence in the regulatory gene; the insertion sequence being selected from IS4, IS10, IS50, IS186, IS231, IS701, IS942, IS1151, IS1170, IS1452 or IS5377, wherein the vehicle is taken up by the host organism, the insertion sequence is inserted in the regulatory gene, and the host organism undergoes sporulation. 2. The composition of claim 1 wherein the host organism comprises Geobacillus stearothermophilus. 3. The composition of claim 1 wherein the reporter gene comprises lacZ, bgaB, xylE, cat, gfp, or a mixture of two or more thereof. 4. The composition of claim 1 wherein the indicator enzyme comprises beta-D-galactosidase, beta-D-glucosidase, alpha-D-glucosidase, alkaline phosphatase, acid phosphatase, butyrate esterase, caprylate esterase lipase, chloroamphenicol acetytransferase, catechol-2,3-dioxygenase, myristate lipase, leucine aminopeptidase, valine aminopeptidase, chymotrypsin, phosphohydrolase, alpha-D-galactosidase, alpha-L-arabinofuranosidase, N-acetyl-beta-glucosaminidase, beta-D-cellobiosidase, alanine aminopeptidase, proline aminopeptidase, tyrosine aminopeptidase, phenylalanine aminopeptidase, beta-D-glucuronidase, fatty acid esterase, or a mixture of two or more thereof. 5. The composition of claim 1 wherein the reporter gene comprises bgaB, the reporter gene producing an indicator enzyme comprising beta-galactosidase. 6. The composition of claim 1 wherein the regulatory gene comprises xylR, lacI, tetR, or a mixture of two or more thereof. 7. The composition of claim 1 wherein the insertion sequence comprises IS5377. 8. The composition of claim 1 wherein the vehicle comprises a plasmid or a viral vector. 9. The composition of claim 8 wherein the plasmid comprises a circular double-stranded DNA. 10. The composition of claim 8 wherein the plasmid is linear. 11. The composition of claim 8 wherein the size of the plasmid is in the range from about 2000 to about 20000 base pairs. 12. The composition of claim 8 wherein from 1 to about 3000 copies of the plasmid are taken up by a cell of the test organism. 13. The composition of claim 8 wherein the plasmid comprises one or more origins of replication. 14. The composition of claim 8 wherein the plasmid comprises one or more genetic markers. 15. The composition of claim 8 wherein the plasmid comprises one or more multiple cloning sites. 16. The composition of claim 8 wherein the plasmid comprises one or more genes that provide a selective marker to induce the test organism to retain the plasmid. 17. The composition of claim 8 wherein the plasmid comprises a gram negative origin of replication and a gram positive origin of replication. 18. The composition of claim 17 wherein the gram negative origin of replication comprises Escherichia coli, and the gram positive origin or replication comprises Geobacillus stearothermophilus, Bacillus atrophaeus, or a mixture thereof. 19. The composition of claim 8 wherein the plasmid comprises an antibiotic resistance gene and/or a gene with exogenous nutritional capability. 20. The composition of claim 8 wherein the plasmid comprises a chloramphenicol, ampicillin or spectinomycin antibiotic gene, and/or a xylose, lactose or amino acid nutritional gene. 21. The composition of claim 8 wherein the viral vector comprises at least one gene transporter that comprises nucleic acid surrounded by a capsid. 22. The composition of claim 8 wherein the viral vector comprises at least one bacteriophage. 23. The composition of claim 8 wherein the viral vector comprises lambda or M13 bacteriophage. 24. The composition of claim 1 wherein the reporter gene comprises bgaB, the regulatory gene comprises xylR, the vehicle is a plasmid, and the host organism comprises Geobacillus stearothermophilus.

Details for Patent 10,011,843

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Emergent Biodefense Operations Lansing Llc	BIOTHRAX	anthrax vaccine adsorbed	Injection	103821	11/12/1998		2034-10-13