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Usefulness of autoantigens depletion to detect autoantibody signatures by multiple affinity protein profiling
Author(s) -
Hardouin Julie,
Lasserre JeanPaul,
Canelle Ludovic,
Duchateau Magalie,
Vlieghe Céline,
ChoquetKastylevsky Geneviève,
JoubertCaron Raymonde,
Caron Michel
Publication year - 2007
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200600324
Subject(s) - autoantibody , antibody , affinity chromatography , antigen , computational biology , chemistry , biology , microbiology and biotechnology , biochemistry , immunology , enzyme
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.