Usefulness of autoantigens depletion to detect autoantibody signatures by multiple affinity protein profiling.
Patients with cancer produce specific autoantibodies against protein antigens present in limited amount among a large background of immunoglobulins (Igs), nonrelevant as biomarkers, including natural antibodies. Multiple affinity protein profiling (MAPPing) that combines 2-D immunoaffinity chromatography, enzymatic digestion of the isolated proteins, and identification by MS/MS, may facilitate the identification of these so far unknown patient antibodies. The first immunoaffinity chromatography is crucial, as it is used for selectively removing proteins (autoantigens) recognized by natural antibodies. Application of this depletion step to colon cancer cell proteins is specifically described along with the identification of the natural autoantigens, as well as the coupling of this depletion step with the next steps. By enabling to separate antibody-binding proteins recognized by either natural autoantibodies or patient-specific antibodies this approach may contribute significantly towards the definition of autoantibody signatures.
Hardouin
Julie
J
Laboratory of Protein Biochemistry and Proteomics, UMR CNRS 7033 (BioMoCeTi), UFR SMBH, University Paris 13, Bobigny cedex, France.
Lasserre
Jean-Paul
JP
Canelle
Ludovic
L
Duchateau
Magalie
M
Vlieghe
Céline
C
Choquet-Kastylevsky
Geneviève
G
Joubert-Caron
Raymonde
R
Caron
Michel
M
D000230:Adenocarcinoma / Q000235:genetics / Q000276:immunology; D000368:Aged; D000595:Amino Acid Sequence; D000912:Antibodies, Neoplasm / Q000302:isolation & purification; D000951:Antigens, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D001323:Autoantibodies / Q000302:isolation & purification*; D001324:Autoantigens / Q000235:genetics / Q000302:isolation & purification*; D018938:Caco-2 Cells; D002846:Chromatography, Affinity / Q000379:methods*; D003110:Colonic Neoplasms / Q000235:genetics / Q000276:immunology; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D040081:Protein Array Analysis / Q000379:methods*; D053719:Tandem Mass Spectrometry
D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D001323:Autoantibodies; D001324:Autoantigens