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Fen1 as marker for chronic obstructive pulmonary disease (copd)

By measuring the protein FEN1 concentration in body fluid samples, it solves the problem in the existing technology that COPD evaluation is expensive and relies on patient cooperation, achieves early diagnosis and the ability to distinguish COPD from other respiratory diseases, and provides an economical and reliable diagnostic method. .

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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  • Fen1 as marker for chronic obstructive pulmonary disease (copd)
  • Fen1 as marker for chronic obstructive pulmonary disease (copd)
  • Fen1 as marker for chronic obstructive pulmonary disease (copd)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0219] Example 1: COPD Study Population

[0220] Source of serum samples:

[0221] 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.

[0222] Serum sample preparation:

[0223]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 t...

Embodiment 21

[0224] Example 2.1: Generation of antibodies against the marker protein FEN1

[0225] Generation of polyclonal antibodies against the marker protein FEN1 for further use of the antibodies to measure serum and plasma levels of FEN1 or concentrations in other bodily fluids by immunodetection assays such as Western blot and ELISA.

[0226] Recombinant protein expression in E. coli:

[0227] To generate antibodies against FEN1, the recombinant antigen was generated in E. coli: therefore, PCR was performed from a full-length cDNA clone (obtained from the German Resource Center for Genome Research (RZPD, Berlin, Germany)) using the following primers Amplification of the FEN1 coding region.

[0228] Forward primer (SEQ ID NO:8):

[0229] 5'-cacaca caattg attaaagaggagaaattaactATGAGAGGATCGCATCACCATCACCATCACATTGAAGGCCGTGGAATTCAAGGCCTGGCC-3' (MunI site is underlined, coding nucleotides are capital letters),

[0230] Reverse primer (SEQ ID NO:9):

[0231] 5'-acgtacgt aagctt TCATTA...

Embodiment 22

[0247] Example 2.2: CRP

[0248] 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 FEN1 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 FEN1 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 the protein FEN1 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 FEN1 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 its characteristic low airflow on...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2333/914G01N33/6884G01N2800/60G01N2800/122G01N2333/916
Inventor J.卡尔J.里德林格N.维尔德
Owner F HOFFMANN LA ROCHE & CO AG
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