Method for identifying immunoreactive peptides

Abstract

The invention relates to a method for identifying immunoreactive peptides. According to said method, a sample of tumorous and corresponding healthy tissue is first provided, the tumor-specific expression profile is subsequently determined and antigenic peptides are isolated from the tumorous tissue and analyzed. The respective data that has been obtained is then matched and peptides are identified on the basis of said data.

1 . Method for identifying immunoreactive peptides comprising the following steps: 
 a) providing a sample of tumorous and corresponding healthy tissue,    b) determining the tumor-specific expression profile in the provided sample,    c) isolation and analysis of antigenic peptides in the sample of tumorous tissue,    d) matching the data obtained in steps b) and c), and    e) identification of peptides on the basis of the matched data.    
     
     
         2 . Method according to  claim 1 , wherein the antigenic peptides are MHC-ligands.  
     
     
         3 . Method according to  claim 1 , wherein step b) is performed by microarray analysis and/or reversed transcriptase-polymerase chain reaction.  
     
     
         4 . Method according to  claim 2 , wherein step b) is performed by microarray analysis and/or reversed transcriptase-polymerase chain reaction.  
     
     
         5 . Method according to  claim 1 , wherein in step c) analysis is performed by mass spectrometry.  
     
     
         6 . Method according to  claim 2 , wherein in step c) analysis is performed by mass spectrometry.  
     
     
         7 . Method according to  claim 3 , wherein in step c) analysis is performed by mass spectrometry.  
     
     
         8 . Method according to  claim 5 , wherein in step c) candidate antigenic peptides are predicted on basis of the expression profile using suitable databases, and that the mass spectrometer is calibrated in view of the peptides.  
     
     
         9 . Method according to  claim 6 , wherein in step c) candidate antigenic peptides are predicted on basis of the expression profile using suitable databases, and that the mass spectrometer is calibrated in view of the peptides.  
     
     
         10 . Method according to  claim 7 , wherein in step c) candidate antigenic peptides are predicted on basis of the expression profile using suitable databases, and that the mass spectrometer is calibrated in view of the peptides.  
     
     
         11 . Method according to  claim 1 , wherein after step c) a further step is performed, in which the reactivity of peripheral leukocytes, preferably T-lymphocytes, is tested against the isolated antigenic peptides.  
     
     
         12 . Method according to  claim 2 , wherein after step c) a further step is performed, in which the reactivity of peripheral leukocytes, preferably T-lymphocytes, is tested against the isolated antigenic peptides.  
     
     
         13 . Method according to  claim 3 , wherein after step c) a further step is performed, in which the reactivity of peripheral leukocytes, preferably T-lymphocytes, is tested against the isolated antigenic peptides.  
     
     
         14 . Method according to  claim 4 , wherein after step c) a further step is performed, in which the reactivity of peripheral leukocytes, preferably T-lymphocytes, is tested against the isolated antigenic peptides.  
     
     
         15 . Method according to  claim 5 , wherein after step c) a further step is performed, in which the reactivity of peripheral leukocytes, preferably T-lymphocytes, is tested against the isolated antigenic peptides.  
     
     
         16 . Method according to  claim 11 , wherein the reactivity test is performed by means of measuring cytokine-mRNA and/or γ-Interferon mRNA synthesized by the leukocytes.  
     
     
         17 . Method according to  claim 1 , wherein after step c) a further step is performed, in which the presence of specific T-lymphocytes is detected.  
     
     
         18 . Method according to  claim 2 , wherein the reactivity test is performed by means of measuring cytokine-mRNA and/or γ-Interferon mRNA synthesized by the leukocytes.  
     
     
         19 . Method according to  claim 3 , wherein the reactivity test is performed by means of measuring cytokine-mRNA and/or γ-Interferon mRNA synthesized by the leukocytes.  
     
     
         20 . Method according to  claim 4 , wherein the reactivity test is performed by means of measuring cytokine-mRNA and/or γ-Interferon mRNA synthesized by the leukocytes.  
     
     
         21 . Method according to  claim 17 , wherein detection of specific T-lymphocytes is carried out by means of labeling leukocytes with reconstituted complexes of antigen-presenting molecules and antigenic peptide.  
     
     
         22 . Method for preparing an immunoreactive peptide, in which a peptide is identified according to the method of  claim 1  and that the identified peptide is synthesized chemically, in vitro or in vivo.  
     
     
         23 . Method for preparing an immunoreactive peptide, in which a peptide is identified according to the method of  claim 2  and that the identified peptide is synthesized chemically, in vitro or in vivo.  
     
     
         24 . Method for preparing an immunoreactive peptide, in which a peptide is identified according to the method of  claim 3  and that the identified peptide is synthesized chemically, in vitro or in vivo.  
     
     
         25 . Method for preparing an immunoreactive peptide, in which a peptide is identified according to the method of  claim 4  and that the identified peptide is synthesized chemically, in vitro or in vivo.  
     
     
         26 . Method for preparing an immunoreactive peptide, in which a peptide is identified according to the method of  claim 5  and that the identified peptide is synthesized chemically, in vitro or in vivo.  
     
     
         27 . Method for preparing an immunoreactive peptide, in which a peptide is identified according to the method of  claim 6  and that the identified peptide is synthesized chemically, in vitro or in vivo.  
     
     
         28 . Method for preparing an immunoreactive peptide, in which a peptide is identified according to the method of  claim 7  and that the identified peptide is synthesized chemically, in vitro or in vivo.  
     
     
         29 . Method for preparing an immunoreactive peptide, in which a peptide is identified according to the method of  claim 8  and that the identified peptide is synthesized chemically, in vitro or in vivo.  
     
     
         30 . Immunoreactive peptide identified by a method according to  claim 1 .  
     
     
         31 . Immunoreactive peptide, said peptide being prepared by a method according to  claim 10 .  
     
     
         32 . Immunoreactive peptide identified by a method according to  claim 2 .  
     
     
         33 . Immunoreactive peptide identified by a method according to  claim 3 .  
     
     
         34 . Immunoreactive peptide identified by a method according to  claim 4 .  
     
     
         35 . Immunoreactive peptide identified by a method according to  claim 5 .  
     
     
         36 . Immunoreactive peptide identified by a method according to  claim 6 .  
     
     
         37 . Immunoreactive peptide identified by a method according to  claim 7 .  
     
     
         38 . Immunoreactive peptide identified by a method according to  claim 8 .  
     
     
         39 . Pharmaceutical composition comprising one or more peptides according to  claim 30 .  
     
     
         40 . Method for treating a tumor disease comprising the step of administering a peptide according to  claim 11 .  
     
     
         41 . Method for assessing the therapy-course of a tumor disease, the method comprising the step of using a peptide according to  claim 30 .  
     
     
         42 . Method for preparing an antibody, wherein a peptide according to  claim 30  is used.  
     
     
         43 . Nucleic acid molecule encoding a peptide according to  claim 30 .  
     
     
         44 . Vector comprising a nucleic acid molecule according to  claim 42 .  
     
     
         45 . Cell genetically modified by a nucleic acid molecule according to  claim 43 , which is producing a peptide according to  claim 30 .  
     
     
         46 . Method for preparing a vaccine comprising the steps of: 
 a) performing a method according to  claim 1;     b) preparing the identified immunoreactive peptides; and    c) formulating the prepared immunoreactive peptides.    
     
     
         47 . Method for preparing a vaccine comprising the steps of: 
 a) performing a method according to  claim 2 , and    b) preparing the identified immunoreactive peptides and    c) formulating the prepared immunoreactive peptides.    
     
     
         48 . Method for preparing a vaccine comprising the steps of: 
 a) performing a method according to  claim 3 , and    b) preparing the identified immunoreactive peptides and    c) formulating the prepared immunoreactive peptides.    
     
     
         49 . Method for preparing a vaccine comprising the steps of: 
 a) performing a method according to  claim 4 , and    b) preparing the identified immunoreactive peptides and    c) formulating the prepared immunoreactive peptides.    
     
     
         50 . Method for preparing a vaccine comprising the steps of: 
 a) performing a method according to  claim 5 , and    b) preparing the identified immunoreactive peptides and    c) formulating the prepared immunoreactive peptides.    
     
     
         51 . Method for preparing a vaccine comprising the steps of: 
 a) performing a method according to  claim 6 , and    b) preparing the identified immunoreactive peptides and    c) formulating the prepared immunoreactive peptides.    
     
     
         52 . Method for preparing a vaccine comprising the steps of: 
 a) performing a method according to  claim 7 , and    b) preparing the identified immunoreactive peptides and    c) formulating the prepared immunoreactive peptides.    
     
     
         53 . Method for preparing a vaccine comprising the steps of: 
 a) performing a method according to  claim 8 , and    b) preparing the identified immunoreactive peptides and    c) formulating the prepared immunoreactive peptides.