IP Library › Granted Patent US 9,206,464
Granted Patent B2
US 9,206,464 · App. 12/749,652 · Granted Dec 8, 2015

Fibrosis biomarker assay

Inventors: Sanne S. Veidal (Olstykke, DK); Morten A. Karsdal (Copenhagen, DK); Diana J. Leeming (Hoor, SE); Natasha Barascuk (Copenhagen O, DK); Helene Skjot-Arkil (Copenhagen S, DK); Antonio Segovia-Silvestre (Copenhagen N, DK); Efstathios Vassiliadis (Rødovre, DK)
Assignee: Nordic Bioscience A/S
C12Q1/37G01N33/6893G01N2800/32
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Quick Facts
Patent No.
US 9,206,464
App. No.
12/749,652
Granted
Dec 8, 2015
Kind
B2
Abstract

Provided herein are methods of diagnosis or of quantitation of fibrosis. An immunoassay is conducted to measure neo-epitope containing protein fragments of collagen type III, collagen type I, collagen type IV, collagen type V, or collagen type VI, elastin, biglycan, decorin, lumican, versican, perlecan, neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, vimentin, or C-reactive protein naturally present in a biofluid sample obtained from a patient. An above normal elevation of the measured protein fragments in the patient is associated with the presence or extent of fibrosis.

Claims (237)

1. A method of diagnosis or of quantitation of fibrosis comprising obtaining a patient biofluid sample, conducting an immunoassay to measure fragments of collagen type VI having an N- or C-terminal neo epitope formed by cleavage of collagen type VI by MMP2, MMP9, MMP13, FAP-1 or FAP-2, said fragments being naturally present in said sample, and associating an elevation of said measure in said patient above a normal level with the presence or extent of fibrosis, wherein said immunoassay is conducted by a method comprising:

contacting the fragments of collagen type VI having said N- or C-terminal neo epitope that are naturally present in said sample with an immunological binding partner specifically reactive with the N- or C-terminal neo-epitope but not reactive with intact collagen type VI, and measuring the extent of binding of the N- or C-terminal neo-epitope to said immunological binding partner to measure therein fragments comprising said neo-epitope,

wherein said immunological binding partner is raised against a synthetic peptide corresponding to an N-terminal neo epitope amino acid sequence formed by cleavage of collagen type VI by MMP2, MMP9, MMP13, FAP-1 or FAP-2 and specifically binds a neo-epitope constituted by said N-terminal amino acid sequence, said N-terminal amino acid sequence selected from the group consisting of:

Collagen type VI

YRGPEG

SEQ ID NO

PIGPKG

SEQ ID NO

GIGIGN

SEQ ID NO

883

865

885

ISGPRG

SEQ ID NO

PGPAGP

SEQ ID NO

VAAKPA

SEQ ID NO

886

887

888

GEPGPP

SEQ ID NO

RGPIGS

SEQ ID NO

PPPPQP

SEQ ID NO

675

889

890

AQGPAG

SEQ ID NO

LIGEQG

SEQ ID NO

PGLIGE

SEQ ID NO

891

892

893

GEPGLN

SEQ ID NO

IGPKGI

SEQ ID NO

VAVVQH

SEQ ID NO

894

895

896

FGPSAA

SEQ ID NO

GPKGET

SEQ ID NO

LGPMGV

SEQ ID NO

897

898

899

PGEPGP

SEQ ID NO

784

or said immunological binding partner is raised against a synthetic peptide corresponding to a C-terminal neo epitope amino acid sequence formed by cleavage of collagen type VI by MMP2, MMP9, MMP13, FAP-1 or FAP-2 and specifically binds a neo-epitope constituted by said C-terminal amino acid sequence, said C-terminal amino acid sequence selected from the group consisting of:

Collagen type VI

GDEGPP

SEQ ID NO

GNADIT

SEQ ID NO

PAGPPG

SEQ ID NO

900

901

133

DPGLMG

SEQ ID NO

PEVPRP

SEQ ID NO

TGPKGI

SEQ ID NO

902

903

904

GDEGGP

SEQ ID NO

PARSAS

SEQ ID NO

TPAPPG

SEQ ID NO

905

906

915

ISGPRG

SEQ ID NO

GISGPR

SEQ ID NO

GIGNRG

SEQ ID NO

886

907

908

YRGYPG

SEQ ID NO

KVEFSL

SEQ ID NO

GVPGRD

SEQ ID NO

909

910

911

PGETGK

SEQ ID NO

RTGPLG

SEQ ID NO

APGERG

SEQ ID NO

912

913

914.

2. A method of immunoassay to measure fragments of collagen type VI having an N- or C-terminal neo epitope formed by cleavage of collagen type VI by MMP2, MMP9, MMP13, FAP-1 or FAP-2, said fragments being naturally present in a biofluid sample, wherein said immunoassay is conducted by a method comprising:

contacting the fragments of collagen type VI having said N- or C-terminal neo epitope that are naturally present in said sample with an immunological binding partner specifically reactive with the N- or C-terminal neo-epitope but not reactive with intact collagen type VI, and measuring the extent of binding of the N- or C-terminal neo-epitope to said immunological binding partner to measure therein fragments comprising said neo-epitope,

wherein said immunological binding partner is raised against a synthetic peptide corresponding to an N-terminal neo epitope amino acid sequence formed by cleavage of collagen type VI by MMP2, MMP9, MMP13, FAP-1 or FAP-2 and specifically binds a neo-epitope constituted by said N-terminal amino acid sequence, said N-terminal amino acid sequence selected from the group consisting of:

Collagen type VI

YRGPEG

SEQ ID NO

PIGPKG

SEQ ID NO

GIGIGN

SEQ ID NO

883

865

885

ISGPRG

SEQ ID NO

PGPAGP

SEQ ID NO

VAAKPA

SEQ ID NO

886

887

888

GEPGPP

SEQ ID NO

RGPIGS

SEQ ID NO

PPPPQP

SEQ ID NO

675

889

890

AQGPAG

SEQ ID NO

LIGEQG

SEQ ID NO

PGLIGE

SEQ ID NO

891

892

893

GEPGLN

SEQ ID NO

IGPKGI

SEQ ID NO

VAVVQH

SEQ ID NO

894

895

896

FGPSAA

SEQ ID NO

GPKGET

SEQ ID NO

LGPMGV

SEQ ID NO

897

898

899

PGEPGP

SEQ ID NO

784

or wherein said immunological binding partner is raised against a synthetic peptide corresponding to a C-terminal neo epitope amino acid sequence formed by cleavage of collagen type VI by MMP2, MMP9, MMP13, FAP-1 or FAP-2 and specifically binds a neo-epitope constituted by said C-terminal amino acid sequence, said C-terminal amino acid sequence selected from the group consisting of:

Collagen type VI

GDEGPP

SEQ ID NO

GNADIT

SEQ ID NO

PAGPPG

SEQ ID NO

900

901

133

DPGLMG

SEQ ID NO

PEVPRP

SEQ ID NO

TGPKGI

SEQ ID NO

902

903

904

GDEGGP

SEQ ID NO

PARSAS

SEQ ID NO

TPAPPG

SEQ ID NO

905

906

915

ISGPRG

SEQ ID NO

GISGPR

SEQ ID NO

GIGNRG

SEQ ID NO

886

907

908

YRGYPG

SEQ ID NO

KVEFSL

SEQ ID NO

GVPGRD

SEQ ID NO

909

910

911

PGETGK

SEQ ID NO

RTGPLG

SEQ ID NO

APGERG

SEQ ID NO

912

913

914.

3. A method of immunoassay to measure fragments of collagen type VI having an N-terminal neo epitope formed by cleavage of collagen type VI by MMP2, said fragments being naturally present in a biofluid sample, wherein said immunoassay is conducted by a method comprising:

contacting the fragments of collagen type VI having said N-terminal neo epitope that are naturally present in said sample with an immunological binding partner specifically reactive with the N-terminal neo-epitope but not reactive with intact collagen type VI, and measuring the extent of binding of the N-terminal neo-epitope to said immunological binding partner to measure therein fragments comprising said neo-epitope,

wherein said immunological binding partner is raised against a synthetic peptide corresponding to an N-terminal neo epitope amino acid sequence formed by cleavage of collagen type VI by MMP2 and specifically binds a neo-epitope constituted by said N-terminal neo epitope amino acid sequence, said N-terminal amino acid sequence being YRGPEG SEQ ID NO883.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 4, 2010
From: VEIDAL, SANNE S.; KARSDAL, MORTEN A.; LEEMING, DIANA J.; BARASCUK, NATASHA; SKJOT-ARKIL, HELENE; SEGOVIA-SILVESTRE, ANTONIO; VASSILIADIS, EFSTATHIOS
To: NORDIC BIOSCIENCE A/S
Reel/Frame 024328/0965 →
Priority Claims (3)
GB 0721713.6 · Nov 5, 2007 · national
GB 0722748.1 · Nov 20, 2007 · national
GB 0802814.4 · Feb 15, 2008 · national
Continuity (4)
Continuation In Part PCTEP2008064946 · Nov 4, 2008
Provisional Application 61211467 · Mar 30, 2009
Provisional Application 61289081 · Dec 22, 2009
Related Publication 20100209940A1 · Aug 19, 2010