Plasma kallikrein fragments as diagnostic biomarkers for lung cancers

Abstract

Disclosed herein are diagnostic markers for lung cancer, isolated from serum glycoproteins. The disclosed diagnostic markers for lung cancer are specifically expressed only in the sera of lung cancer patients at high levels, and thus will be very useful for diagnosing lung cancer and estimating disease progression and treatment.

1 . A diagnostic marker for lung cancer, which comprises a protein selected from the group consisting of complement component c8 beta chain precursor, protein s100-a9, C4b-binding protein alpha chain precursor, isoform 2 of apolipoprotein-11 precursor, protein s100-a8, proteins similar to cavia porcellus phosphatidic acid phosphatase 2A mRNA, complement component c9 precursor, inter-alpha-trypsin inhibitor heavy chain h3 precursor, F1j00385 protein (fragment), coagulation factor xii precursor, Ig alpha-2 chain c region, complement component c8 alpha chain precursor, corticosteroid-binding globulin precursor, clusterin precursor, gamma-g globin, serum amyloid p-component precursor, plasma kallikrein (KLKB1) fragment, Igha1 protein, heparin cofactor 2 precursor, 16 kDa protein, Ig kappa chain v-iii region go1, complement factor I precursor, hemoglobin subunit beta, proteins similar to tripartite motif protein 49, isoform 1 of complement factor b precursor (fragment), alpha-1-antitrypsin precursor, carboxypeptidase n subunit 2 precursor, complement factor h-related protein 3 precursor, plasma protease c1 inhibitor precursor, Ig kappa chain c region, alpha 2 macroglobulin variant, dermcidin precursor, complement c5 precursor and pregnancy zone protein precursor. 
     
     
         2 . The diagnostic marker of  claim 1 , which comprises inter-alpha-trypsin inhibitor heavy chain h3 precursor. 
     
     
         3 . The diagnostic marker of  claim 1 , which comprises plasma kallikrein (KLKB1) fragment. 
     
     
         4 . A composition for diagnosing lung cancer, which comprises an antibody binding specifically to a protein selected from the group consisting of complement component c8 beta chain precursor, protein s100-a9, C4b-binding protein alpha chain precursor, isoform 2 of apolipoprotein-11 precursor, protein s100-a8, proteins similar to cavia porcellus phosphatidic acid phosphatase 2A mRNA, complement component c9 precursor, inter-alpha-trypsin inhibitor heavy chain h3 precursor, F1j00385 protein (fragment), coagulation factor xii precursor, Ig alpha-2 chain c region, complement component c8 alpha chain precursor, corticosteroid-binding globulin precursor, clusterin precursor, gamma-g globin, serum amyloid p-component precursor, plasma kallikrein (KLKB1) fragment, Igha1 protein, heparin cofactor 2 precursor, 16 kDa protein, Ig kappa chain v-iii region go1, complement factor I precursor, hemoglobin subunit beta, proteins similar to tripartite motif protein 49, isoform 1 of complement factor b precursor (fragment), alpha-1-antitrypsin precursor, carboxypeptidase n subunit 2 precursor, complement factor h-related protein 3 precursor, plasma protease c1 inhibitor precursor, Ig kappa chain c region, alpha 2 macroglobulin variant, dermcidin precursor, complement c5 precursor and pregnancy zone protein precursor. 
     
     
         5 . The composition of  claim 4 , which comprises an antibody binding specifically to inter-alpha-trypsin inhibitor heavy chain h3 precursor. 
     
     
         6 . The composition of  claim 4 , which comprises an antibody binding specifically to plasma kallikrein (KLKB1) fragment. 
     
     
         7 . A kit for diagnosing lung cancer, which comprises an antibody binding specifically to inter-alpha-trypsin inhibitor heavy chain h3 precursor. 
     
     
         8 . A protein chip for diagnosing lung cancer, which has attached thereto an antibody binding specifically to inter-alpha-trypsin inhibitor heavy chain h3 precursor. 
     
     
         9 . A kit for diagnosing lung cancer, which comprises an antibody binding specifically to plasma kallikrein (KLKB1) fragment. 
     
     
         10 . A protein chip for diagnosing lung cancer, which has attached thereto an antibody binding specifically to plasma kallikrein (KLKB1) fragment. 
     
     
         11 . A composition for diagnosing lung cancer, which comprises a primer or probe specific for a nucleic acid encoding a protein selected from the group consisting of complement component c8 beta chain precursor, protein s100-a9, C4b-binding protein alpha chain precursor, isoform 2 of apolipoprotein-11 precursor, protein s100-a8, proteins similar to cavia porcellus phosphatidic acid phosphatase 2A mRNA, complement component c9 precursor, inter-alpha-trypsin inhibitor heavy chain h3 precursor, F1j00385 protein (fragment), coagulation factor xii precursor, Ig alpha-2 chain c region, complement component c8 alpha chain precursor, corticosteroid-binding globulin precursor, clusterin precursor, gamma-g globin, serum amyloid p-component precursor, plasma kallikrein (KLKB1) fragment, Igha1 protein, heparin cofactor 2 precursor, 16 kDa protein, Ig kappa chain v-iii region go1, complement factor I precursor, hemoglobin subunit beta, proteins similar to tripartite motif protein 49, isoform 1 of complement factor b precursor (fragment), alpha-1-antitrypsin precursor, carboxypeptidase n subunit 2 precursor, complement factor h-related protein 3 precursor, plasma protease c1 inhibitor precursor, Ig kappa chain c region, alpha 2 macroglobulin variant, dermcidin precursor, complement c5 precursor and pregnancy zone protein precursor. 
     
     
         12 . The composition of  claim 11 , which comprises a primer or probe specific for a nucleic acid encoding inter-alpha-trypsin inhibitor heavy chain h3 precursor (ITI-H3). 
     
     
         13 . The composition of  claim 11 , which comprises a primer or probe specific for a nucleic acid encoding plasma kallikrein fragment. 
     
     
         14 . A kit for diagnosing lung cancer, which comprises a primer or probe specific for a nucleic acid encoding inter-alpha-trypsin inhibitor heavy chain h3 precursor. 
     
     
         15 . A DNA chip for diagnosing lung cancer, which has attached thereto a primer or probe specific for a nucleic acid encoding inter-alpha-trypsin inhibitor heavy chain h3 precursor. 
     
     
         16 . A kit for diagnosing lung cancer, which comprises a primer or probe specific for a nucleic acid encoding plasma kallikrein fragment. 
     
     
         17 . A DNA chip for diagnosing lung cancer, which has attached thereto a primer or probe specific for a nucleic acid encoding plasma kallikrein fragment. 
     
     
         18 . A method for diagnosing lung cancer, which comprises the steps of:
 (a) bringing a biological sample into contact with an antibody binding specifically to the diagnostic marker of  claim 1 ; and   (b) detecting the formation of an antigen-antibody complex.   
     
     
         19 . method of  claim 18 , wherein the biological sample in the step (a) is serum. 
     
     
         20 . method of  claim 18 , wherein the detection of the formation of the antigen-antibody in the step (b) is carried out using a method selected from the group consisting of RIA (radioimmunoassay), RIPA (radioimmunoprecipitation assay), IFA (immunoflourescence assay), EILSA (enzyme-linked immunosorbent assay) and Western blot. 
     
     
         21 . A method for diagnosing lung cancer, which comprises the steps of:
 (a) removing a plasma kallikrein fragment and the heavy chain precursor form thereof from the serum of a subject;   (b) bringing the serum, from which the plasma kallikrein fragment and the heavy chain precursor form thereof had been removed in the step (a), into contact with an antibody specific for a region comprising the H4 domain of the plasma kallikrein fragment; and   (c) detecting the formation of an antigen-antibody complex.   
     
     
         22 . The method of  claim 21 , wherein the removal of the plasma kallikrein fragment or the heavy chain precursor form thereof in the step (a) is carried out by bringing the serum into contact with an antibody binding specifically to the plasma kallikrein fragment or is carried out using a molecular size separation column. 
     
     
         23 . method of  claim 21 , wherein the detection of the antigen-antibody complex in the step (c) is carried out using a method selected from the group consisting of RIA (radioimmunoassay), RIPA (radioimmunoprecipitation assay), IFA (immunoflourescence assay), EILSA (enzyme-linked immunosorbent assay) and Western blot. 
     
     
         24 . method of  claim 21 , wherein, if a fragment of 15-20 kDa is detected in the step (c), the subject is diagnosed to have lung cancer. 
     
     
         25 . A method for identifying diagnostic markers for lung caner, which comprises the steps of:
 (a) separating glycoproteins from a serum sample using a multi-lectin affinity column;   (b) concentrating the separated glycoproteins of step (a) by acetone precipitation;   (c) deglycosylating the concentrated glycoproteins of step (b) by enzymatic treatment;   (d) subjecting the deglycosylated proteins of step (c) to SDS-PAGE;   (e) obtaining peptides from the proteins by in-gel trypsin digestion; and   (f) analyzing the peptides of step (e) by LC-MS/MS to identify proteins which are specifically expressed only in the sera of lung cancer patients compared to normal sera.