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Exploring cDNA Phage Display for Autoantibody Profiling in the Serum of Multiple Sclerosis Patients
Author(s) -
GOVARTS C.,
SOMERS K.,
HUPPERTS R.,
STINISSEN P.,
SOMERS V.
Publication year - 2007
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1398.043
Subject(s) - phage display , complementary dna , autoantibody , cdna library , antigen , antibody , selection (genetic algorithm) , peptide library , biology , serology , microbiology and biotechnology , computational biology , immunology , genetics , gene , peptide sequence , computer science , artificial intelligence
: We applied a cDNA phage display method called serological antigen selection (SAS) to identify immunogenic targets that evoke an autoantibody response in the serum of multiple sclerosis (MS) patients. This method involves the display of a cDNA expression library, in this study a MS brain library, on filamentous phage and subsequent selection using patient immunoglobulin G (IgG). To apply the SAS technology for autoantibodies in the serum of MS patients, an optimization was necessary to deplete cDNA products that encode IgG fragments derived from B cells present in the MS brain plaques. We describe a differential screening procedure in which positive selection rounds on MS serum and negative selection rounds on healthy control serum were alternated to optimize the selection procedure. As a result, a substantial decrease of IgG‐displaying phage clones was observed after each negative selection round, thereby preventing an overgrowth of IgG‐displaying phage clones. Our depletion strategy was therefore successful in preventing the enrichment of IgG‐displaying phage clones. This approach will facilitate the identification of possible MS‐related antigens.