Abstract:
The present invention relates to a qualitative and quantitative analytical method for analyzing the activity type of an enzyme that is activated by proteolysis. The method of the present invention is advantageous in that the activity type of an enzyme present in a specimen is analyzed using an enzyme activity inhibitor and a binder to enable the analysis to be performed in a quicker and more accurate manner as compared to enzyme-activity analysis methods using a simple binder (for example, an antibody). The method of the present invention can enable the analysis of the activity type of an enzyme present in a specimen in a simple manner using less equipment than enzyme-activity analysis methods using a simple binder. The method of the present invention involves simultaneously using the inhibitor and the binder in analyzing the activity type of a target enzyme or an enzyme to enable quick and specific analysis, and high-throughput drug screening can be performed at a high speed due to the above-described characteristics of the method of the present invention. In addition, the method of the present invention can be applied to point-of-care testing (POCT) for the clinical monitoring of the effects and dosage of a drug in a drug administration target (for example, an animal or human).
Abstract:
The invention provides nucleic acid molecules, e.g., RNA molecules, that catalyze their own replication and undergo exponential amplification at a constant temperature and in the absence of proteins or other biological components, and methods of preparing and using those molecules.
Abstract:
The subject of the invention is an in cellulo method for screening for agents that modulate the activity of the Mdm2 ubiquitin ligase, said method comprising the following steps: a) bringing a candidate test agent into contact with recombinant yeast cells which express, in the nucleus thereof, a fusion protein comprising: (i) a polypeptide MDM2 and (ii) at least one detectable protein; b) quantifying said first detectable protein in the yeast cells, at the end of at least one predetermined period of time after the candidate agent has been brought into contact with said cells; c) comparing the value obtained in step (b) with a control value obtained when step (a) is carried out in the absence of the candidate agent.
Abstract:
Methods of screening for modulators of TRIM24 (also known as TIF1 -ALPHA) expression and/or biological activity are described. In particular, methods of screening of screening for modulators of TRIM24 E3 ligase activity, and specifically an E3 ligase activity directed at p53 as the target polypeptide are also described. Modulators of TRIM24 expression and activity are provided and their use in treatment of cancer, particularly in breast, colon, prostate, renal cancers and in acute lymphoblastic leukaemia. Suitable modulators of TRIM24 expression include siRNA and shRNA and can be used in the treatment of cancer and for targeting cancer stem cells.
Abstract:
The present invention relates to a prostaglandin D 2 metabolite, derivatives thereof, compositions comprising the metabolite, and an antibody that specically binds to the metabolite. Methods of use are also provided.
Abstract:
NAD-dependent ligase is identified as an indicator of micro¬ organisms in a sample. A method of detecting the presence of an NAD-dependent ligase expressing micro-organism in a sample comprises the steps of: (a) contacting the sample with a nucleic acid molecule which acts as a substrate for NAD-dependent ligase activity in the sample, (b) incubating the thus contacted sample under conditions suitable for NAD-dependent ligase activity; and (c) specifically determining the presence of a ligated nucleic acid molecule resulting from the action of the NAD- dependent ligase on the substrate nucleic acid molecule to indicate the presence of the NAD-dependent ligase expressing micro-organism. The method has a number of applications and kits for carrying out the methods are also provided. Lysostaphin preparations substantially free from nuclease and/or ligase contaminants are produced by heating a lysostaphin preparation which contains nuclease and/or ligase contaminants under conditions whereby nuclease and/or ligase activity is reduced whereas endopeptidase activity of the lysostaphin is substantially unaffected.
Abstract:
The present invention relates to an enzyme activity assay using rolling circle 5 amplification for verifying that a sample contains enzyme activity. The enzyme activity assayed is typically involved in processing of mismatched nucleotides and/or damaged nucleotides in a double stranded nucleic acid. The present invention relates to methods for determining the presence of enzyme activities involved in processing double stranded oligonucleotide. Methods are also directed against determining the presence 10 of nucleotide repair enzyme activities involved in the repair of a circular oligonucleotide. The present invention also relates to liquid compositions and solid support both comprising an oligonucleotide probe. Furthermore, the present invention relates to methods for testing the efficacy of a drug, for diagnosing, prognosing, treating a disease by determining the enzyme activity.
Abstract:
The invention provides an isolated, purified population of human cells comprising CD8+ T cells with reduced Cbl-b activity. The invention provides uses of such cells in methods for inducing or enhancing an anti-tumor immune response in a subject. These methods comprise: (a) providing a cell population, from a subject or from another source, which comprises CD8+ T cells, (b) reducing Cbl-b activity in the CD8+ T-cells, (c) administering the cells of step (b) to the subject. The invention provides methods for making CD8+ T cells that do not require stimulation through a co-receptor in order for the cell to become activated or proliferated in response to contact via its T cell receptor. Such methods are based upon reducing function of Cbl-b. The invention also provides methods for identifying agents which affect Cbl-b expression or activity.