Use of a Fibrinogen Capture Agent to Detect a Ciz1 B-Variant

Abstract

The present invention relates to the use of a fibrinogen capture agent or a fibrinogen detection agent in an assay to detect a Ciz1 b-variant.

1 - 7 . (canceled) 
     
     
         8 . A method of detecting cancer in a subject which comprises detecting a Ciz1 b-variant peptide in a sample from the subject, wherein the presence of Ciz1 b-variant peptide in the sample indicates that the subject has cancer and wherein the method comprises the step of capturing a fibrinogen/Ciz1 b-variant peptide complex from the sample. 
     
     
         9 . A method of detecting cancer in a subject which comprises detecting a Ciz1 b-variant peptide in a sample from the subject, wherein the presence of Ciz1 b-variant peptide in the sample indicates that the subject has cancer and wherein the Ciz1 b-variant peptide is the sequence shown as SEQ ID NO: 11 or a fragment thereof which comprises EVR as shown in positions 36 to 38 of SEQ ID NO: 11. 
     
     
         10 . A method according to  claim 8  which comprises the following steps:
 (a) capturing a fibrinogen/Ciz1 b-variant complex using a fibrinogen capture agent; and 
 (b) detecting Ciz1 b-variant using a Ciz1 b-variant detection agent; or 
 (a) capturing a fibrinogen/Ciz1 b-variant complex using a Ciz1 b-variant capture agent; and 
 (b) detecting fibrinogen using a fibrinogen detection agent. 
 
     
     
         11 . The method according to  claim 10  wherein the Ciz1 b-variant capture agent is an antibody or a fragment thereof, or a non-protein scaffold which specifically binds Ciz1 b-variant. 
     
     
         12 . The method according to  claim 10  wherein the Ciz1 b-variant capture agent is an antibody which specifically binds Ciz1 b-variant. 
     
     
         13 . The method according to  claim 8  wherein the step of detecting the Ciz1 b-variant peptide or fibrinogen employs antibody-based arrays, enzyme linked immunosorbent assays (ELISA), non-antibody protein scaffolds, radioimmuno-assay (RIA), western blotting, aptamers or mass spectrometry. 
     
     
         14 . The method according to  claim 8 , further comprising the step of releasing a Ciz1 b-variant peptide and/or a fibrinogen/Ciz1 b-variant complex from an exosomal compartment of the sample. 
     
     
         15 . The method according to  claim 14  wherein the Ciz1 b-variant peptide and/or fibrinogen/Ciz1 b-variant complex is released from the exosomal compartment by detergent treatment. 
     
     
         16 . The method according to  claim 14 , further comprising the step of enriching the exosomal fraction from the sample. 
     
     
         17 . The method according to  claim 16 , wherein the step of enriching the exosomal fraction comprises ultracentrifugation, ultrafiltration, continuous flow electrophoresis, chromatography, precipitation or cross-flow ultrafiltration. 
     
     
         18 . The method according to  claim 10  wherein the cancer is selected from lung, lymphoma, kidney, breast, liver, bladder, ovarian or thyroid cancer. 
     
     
         19 . The method according to  claim 18  wherein the cancer is lung cancer. 
     
     
         20 . A method of treating a subject with cancer comprising administering to the subject a cancer therapeutic; wherein the subject has been identified as having cancer by the method as defined in  claim 10 . 
     
     
         21 - 23 . (canceled) 
     
     
         24 . A method of detecting a Ciz1 b-variant in a sample which comprises the step of releasing a Ciz1 b-variant and/or a fibrinogen/Ciz1 b-variant complex from an exosomal compartment of the sample by treating the sample with detergent and detecting if the Ciz1 b-variant is present in the sample. 
     
     
         25 . The method according to  claim 24  wherein the Ciz1 b-variant is in a complex with fibrinogen. 
     
     
         26 . The method according to  claim 24  which comprises the step of enriching the exosomal fraction from the sample prior to releasing the Ciz1 b-variant. 
     
     
         27 . The method according to  claim 26 , wherein the step of enriching the exosomal fraction comprises ultracentrifugation, ultrafiltration, continuous flow electrophoresis, chromatography, precipitation or cross-flow ultrafiltration. 
     
     
         28 . A kit comprising an agent which specifically binds fibrinogen and an agent which specifically binds Ciz1 b-variant. 
     
     
         29 . A kit according to  claim 28  wherein the fibrinogen is fibrinogen alpha chain. 
     
     
         30 . A kit according to  claim 28  wherein the agent which specifically binds fibrinogen and/or the agent which specifically binds Ciz1 b-variant is an antibody or a fragment thereof, a non-protein scaffold or an aptamer.