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Assays for Cancer Patient Monitoring Based on Levels of Analyte Components of the Plasminogen Activator System in Body Fluid Samples

a technology of plasminogen activator and cancer patient, which is applied in the direction of material analysis, drug composition, instruments, etc., to achieve the effects of reducing tumor size, avoiding patient subjection, and increasing the efficacy of treatmen

Inactive Publication Date: 2009-11-19
SIEMENS HEALTHCARE DIAGNOSTICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides methods for analyzing body fluid samples from cancer patients to detect and measure the levels of certain proteins, namely uPA, PAI-1, and the complex of uPA:PAI-1, to determine if the levels of these proteins are increased in cancer patients compared to normal individuals. This allows for the monitoring of a cancer patient's disease status and response to therapy. The invention also provides normal plasma or serum values for uPA, PAI-1, and the uPA:PAI-1 complex in healthy individuals. The methods can be used to select cancer patients who are most likely to benefit from anti-cancer or anti-neoplastic disease therapies.

Problems solved by technology

Because the PA system components are intricately involved in the process of cancer and cancer spread in a variety of cancer types, which afflict both genders, it is a problem in the art to be able to accurately and sensitively screen over time to determine and monitor those individuals who are likely to respond, and / or who are responding to, (or not responding to), or benefiting from (or not benefiting from), anti-cancer therapy(ies), or combination therapies, particularly, molecularly targeted therapies to the plasminogen activation system.

Method used

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  • Assays for Cancer Patient Monitoring Based on Levels of Analyte Components of the Plasminogen Activator System in Body Fluid Samples
  • Assays for Cancer Patient Monitoring Based on Levels of Analyte Components of the Plasminogen Activator System in Body Fluid Samples
  • Assays for Cancer Patient Monitoring Based on Levels of Analyte Components of the Plasminogen Activator System in Body Fluid Samples

Examples

Experimental program
Comparison scheme
Effect test

example 1

Enzyme Linked Immunoassay (ELISA) to Measure Levels of uPA in a Body Fluid Sample

[0077]Serum and plasma samples were centrifuged in a tabletop microcentrifuge to remove all flocculent material and / or particulate matter. After centrifugation, the samples were analyzed without further treatment, or were stored at −70° C. for future analysis. The initial concentration of the serum sample examined did not exceed a concentration of 10% (i.e., a 1:10 dilution of sample in sample diluent), (Bayer Diagnostics / Oncogene Science uPA Microtiter ELISA assay kit manual).

[0078]The Bayer Diagnostics / Oncogene Science uPA Microtiter ELISA detects 25 pg / ml of uPA analyte in a test sample. The signal of the 25 pg / ml standard is greater than two times the zero (background) signal. In addition, the uPA ELISA is specific for the detection of uPA; for example, no cross-reactivity is detected against tPA and cathepsin D, even at high challenge doses.

Detailed Protocol

[0079]The uPA Microtiter ELISA kit (comme...

example 2

Enzyme Linked Immunoassay (ELISA) to Measure Levels of PAI-1 in Plasma

[0089]Plasma samples were centrifuged in a tabletop microcentrifuge to remove all flocculent material and / or particulate matter. Plasma samples were typically diluted 1:50 in sample diluent as described below and in accordance with the instructions accompanying the Bayer Diagnostics / Oncogene Science PAI-1 Microtiter ELISA assay. The contents of the PAI-1 Microtiter ELISA assay manual (Bayer Diagnostics / Oncogene Science, Cambridge, Mass.) are hereby incorporated by reference herein in their entirety.

[0090]The Bayer Diagnostics / Oncogene Science PAI-1 Microtiter ELISA detects 0.10 ng / ml of PAI-1 analyte in a test sample. The signal of the 0.10 ng / ml standard is greater than two times the zero (background) signal. In addition, the PAI-1 Microtiter ELISA is capable of quantifying PAI-1 in the active and latent forms, as well as in complex with uPA or tPA. The assay has been tested for cross-reactivity by challenging wi...

example 3

Enzyme Linked Immunoassay (ELISA) to Measure Levels of the uPA:PAI-1 Complex in Plasma

[0101]The Bayer Diagnostics / Oncogene Science Human uPA:PAI-1 Complex Microtiter ELISA detects 5 pg / ml of uPA:PAI-1 complex in a test sample. The signal of the 5 pg / ml standard is significantly higher than the zero (background) signal. In addition, the uPA:PAI-1 complex ELISA assay has been tested for specificity against PAI-2, tPA, ovalbumin, free uPA and free PAI-1 and showed no cross-reactivity against these proteins at high challenge doses.

Detailed Protocol

[0102]The uPA:PAI-1 complex Microtiter ELISA kit (commercially available from Bayer Diagnostics / Oncogene Science, Cambridge, Mass.) used in this Example has removable strips of wells so that the assay can be performed on multiple occasions, and only the number of wells needed are used. A standard curve was included each time samples were analyzed. The standard curve performed for each separate assay requires 12 wells (6 standards run in duplic...

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Abstract

The present invention describes clinically and medically important methods of examining, screening over time, and monitoring the outcome of a cancer patient who is undergoing treatment or therapy for his or her disease. More specifically, the invention provides a method of monitoring the progression of disease, or the effectiveness of cancer treatment, in a cancer patient by measuring the levels of one or more analytes of the plasminogen activator (uPA) system, namely, uPA, PAI-1 and the complex of uPA:PAI-1, in a sample taken from the cancer patient, preferably, before treatment, at the start of treatment, and at various time intervals during treatment. As a result of performing the method, an increase or elevation in the levels of one or more of the PA system analytes in the cancer patient compared with the levels one or more of the respective PA system analytes in normal control individuals serves as an indicator of cancer advancement or progression and / or a lack of treatment effectiveness for the patient.

Description

FIELD OF THE INVENTION[0001]The present invention relates to assays for monitoring or assessing the progress of cancer patients during a course of disease or disease treatment or therapy by determining levels of one or more cancer analytes, i.e., components of the plasminogen activator (PA) system, compared to the levels of one or more of these PA system components in normal control individuals. According to the methods described herein, the determination of increases in the levels of one or more of urokinase plasminogen activator (uPA), PA inhibitor-1 (PAI-1), or a complex of uPA and PAI-1 (the uPA:PAI-1 complex), compared with the levels of these respective analytes in normal controls is indicative of poor patient and / or treatment outcome relative to disease status.BACKGROUND OF THE INVENTION[0002]The plasminogen activator (PA) system involves the serine proteases plasmin and urokinase plasminogen activator (uPA); the serpins α2-antiplasmin, plasminogen activator inhibitor type-1 ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/53G01N33/574G01N33/86
CPCG01N33/57484G01N33/86Y10T436/25G01N2333/9723G01N2800/52G01N2333/8132A61P35/00
Inventor CARNEY, WALTER P.HAMER, PETER J.
Owner SIEMENS HEALTHCARE DIAGNOSTICS INC
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