Seprase as marker for chronic obstructive pulmonary disease (copd)

A chronic obstructive, surface-expressed technology that can be used in disease diagnosis, biological testing, biomaterial analysis, etc., and can solve the problems of time-consuming general and widespread use burden, expensive spirometry, etc.

Inactive Publication Date: 2013-11-27
F HOFFMANN LA ROCHE & CO AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Spirometry is very expensive, time consuming and cannot be afforded by health care systems for general and widespread use in screening large numbers of subjects

Method used

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  • Seprase as marker for chronic obstructive pulmonary disease (copd)
  • Seprase as marker for chronic obstructive pulmonary disease (copd)
  • Seprase as marker for chronic obstructive pulmonary disease (copd)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0220] Example 1: COPD Study Population

[0221] Source of serum samples:

[0222] To identify COPD-specific proteins as potential diagnostic markers of COPD, serum samples were derived from well-characterized COPD patients (according to the ATS classification system in Table 1) in a national multicenter study. From each sample donor, spirometry was performed. Pulmonary function, other diagnostic tests, and cause, diagnosis, and comorbidities of transferal were documented in a dedicated case report form (CRF). COPD samples have been evaluated in comparison to control samples obtained from Control Groups 1-4, as shown in Table 2.

[0223] Serum sample preparation:

[0224] Serum samples were collected into serum tubes and allowed to clot for at least 60 minutes and up to 120 minutes at room temperature. After centrifugation (10 min, 2000 g), the supernatant was divided into 1 ml aliquots and frozen at -70°C. Samples were thawed, aliquoted into smaller volumes suitable for ...

Embodiment 21

[0225] Example 2.1: Antibodies against the marker protein Seprase

[0226] As Seprase-specific binding molecules, two different rat monoclonal anti-Seprase antibodies (clones D8 and D28) were used (Pineiro-Sanchez, M.L. et al., J. Biol. Chem. 12 (1997) 7595-7601).

[0227] Biotinylation of Monoclonal Rat IgG:

[0228] Monoclonal rat IgG (clone D28) in 10 mM NaH 2 PO 4 / NaOH pH7.5, 30mM NaCl to adjust to 10mg / ml. Add 50 μl biotin-N-hydroxysuccinimide (3.6 mg / ml in DMSO) per ml IgG solution. After 30 minutes at room temperature, the sample was placed in Superdex200 (10mM NaH 2 PO 4 / NaOH pH7.5, 30mM NaCl) on the chromatography. Fractions containing biotinylated IgG were pooled. Polyclonal antibodies have been biotinylated following the same protocol.

[0229] Digoxigenation of Monoclonal Rat IgG:

[0230] Monoclonal rat IgG (clone D8) in 10 mM NaH 2 PO 4 / NaOH, 30mM NaCl pH7.5 to adjust to 10mg / ml. Add 50 μl of Digoxigenin-3-O-methylcarbonyl-ε-aminocaproic acid-N-hyd...

Embodiment 22

[0231] Example 2.2: CRP

[0232] The marker protein CRP was measured using a homogenous assay (Hitachi) distributed by Roche Diagnostics (Mannheim, FRG).

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Abstract

The present invention relates to an in vitro method aiding in the assessment of chronic obstructive pulmonary disease (= COPD). It discloses the use of the protein Seprase as a marker of COPD. Furthermore, it especially relates to a method for assessing COPD from a sample, derived from an individual by measuring the protein Seprase in said sample in vitro.

Description

field of invention [0001] The present invention relates to an in vitro method to aid in the assessment of Chronic Obstructive Pulmonary Disease (=COPD). It discloses the use of a protein surface expressed protease (Seprase) as a marker for COPD. Furthermore, it relates in particular to a method for assessing COPD from a sample derived from an individual by measuring the protein Seprase in said sample in vitro. Background of the invention [0002] Chronic obstructive pulmonary disease (COPD) is a disease characterized by chronic inflammation and irreversible airflow obstruction with a faster than normal decline in the lung function parameter FEV1. This results in restricted airflow to and from the lungs, causing shortness of breath. The disease has two main pathological aspects, chronic bronchitis (characterized by hypersecretion of mucus from the guiding airways) and emphysema (characterized by destructive changes in the alveoli). In clinical practice, COPD is defined by ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N2333/96433G01N33/6893G01N33/6884G01N2800/60G01N2800/122
Inventor J.卡尔J.雷德林格W.罗琳格
Owner F HOFFMANN LA ROCHE & CO AG
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