Abstract: Micro-organisms which over-produce ATPase, especially mycoplasmas in a sample of a cell culture, are detected by making use of the ATPase activity to convert cellular or externally added ATP to ADP. The decrease in ATP externally added or the increase or absolute level of ADP produced from cellular ATP can then be detected or measured. For example, if a reagent such as phosphenol pyruvate/pyruvate kinase is added to convert that ADP back to ATP, the resulting high level of ATP can be detected bioluminescently using the reaction: Luciferin+luciferase enzyme+ATP?hv+products.
Type:
Grant
Filed:
May 7, 2002
Date of Patent:
October 4, 2005
Assignee:
Lumitech (UK) Limited
Inventors:
Sharon Patricia Mary Crouch, Dawn Ann Bradbury, Kevin John Slater
Abstract: A method for measuring protein kinase activity comprising: (a) providing a first solution comprising ATP and a protein kinase to be tested, and a second solution comprising ATP in the absence of said kinase to be tested; (b) adding a substrate capable of being phosphorylated by the protein kinase to be tested to the first and second solutions of step (a); (c) measuring the concentration of ATP and/or ADP, or the rate of change thereof with respect to time, in each of the reaction mixtures formed in step (b) using a bioluminescence reaction; and (d) using the information about the concentration of ATP and/or ADP to determine the activity of the protein kinase to be tested.
Type:
Grant
Filed:
May 1, 2003
Date of Patent:
June 28, 2005
Assignee:
Lumitech (UK) Limited
Inventors:
Sharon Patricia Mary Crouch, Kevin John Slater
Abstract: A method for measuring protein kinase activity comprising:
(a) providing a first solution comprising ATP and a protein kinase to be tested, and a second solution comprising ATP in the absence of said kinase to be tested;
(b) adding a substrate capable of being phosphorylated by the protein kinase to be tested to the first and second solutions of step (a);
(c) measuring the concentration of ATP and/or ADP, or the rate of change thereof with respect to time, in each of the reaction mixtures formed in step (b) using a bioluminescence reaction; and
(d) using the information about the concentration of ATP and/or ADP to determine the activity of the protein kinase to be tested.
Type:
Grant
Filed:
December 14, 2001
Date of Patent:
July 29, 2003
Assignee:
LumiTech (UK) Limited
Inventors:
Sharon Patricia Mary Crouch, Kevin John Slater
Abstract: Micro-organisms which over-produce ATPase, especially mycoplasmas in a sample of a cell culture, are detected by making use of the ATPase activity to convert cellular or externally added ATP to ADP. The decrease in ATP externally added or the increase or absolute level of ADP produced from cellular ATP can then be detected or measured. For example, if a reagent such as phosphenol pyruvate/pyruvate kinase is added to convert that ADP back to ATP, the resulting high level of ATP can be detected bioluminescently using the reaction: Luciferin+luciferase enzyme+ATP→&ngr;+products.
Type:
Application
Filed:
May 7, 2002
Publication date:
March 13, 2003
Applicant:
LUMITECH (UK) LIMITED
Inventors:
Sharon Patricia Mary Crouch, Dawn Ann Bradbury, Kevin John Slater
Abstract: The invention relates to methods for detecting the cytotoxic activity of an effector, such as a cytotoxic T lymphocyte, on a non-microbial target cell. Detection of cytotoxic activity is preferably achieved by detecting adenylate kinase release using photometric methods.
Type:
Application
Filed:
December 7, 2001
Publication date:
January 9, 2003
Applicant:
LumiTech (UK) Limited
Inventors:
Sharon Patricia Mary Crouch, Kevin John Slater