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MS\/MS peptide sequencing was performed in conjunction with a micro reversed phase HPLC configured with an online MALDI plate\u2010spotting device. Apheresis treatment had been performed in three patients with longstanding therapy refractory rheumatoid arthritis. 2\u2010D gels displayed <jats:italic>ca.<\/jats:italic> 500 spots representing proteins that were eluted from the Prosorba<jats:sup>\u00ae<\/jats:sup> columns. From 54 gels, a total of 1256 protein spots had been picked and yielded in the identification of 56 non\u2010redundant proteins without counting isoforms. Proteins from the eluates belong to five major groups comprising (i) immunoglobulins (IgG, IgA, IgM heavy and light chains; about 40% of the spots), (ii) proteins involved in coagulation, (iii) HDL\/LDL\u2010associated proteins, (iv) proteins from the complement system, and (v) acute phase proteins. MS analysis showed that the full\u2010length C3 complement protein had been cleaved upon complement activation, presumably on the column, such that the anaphylatoxin C3a was produced and released during therapy. Our results are consistent with clinical observations on both patient responses to therapy and reported adverse events. For the first time, direct molecular information has become available to support mechanistic reasoning for the principle of function of staphylococcal protein A\u2010based immunoadsorption therapy and for the explanation of adverse events. According to our results, removal and\/or modulation of immune complexes together with complement activation can be regarded as the major events that are taking place during Prosorba<jats:sup>\u00ae<\/jats:sup> therapy. In order to avoid complement activation and induction of an inflammatory cascade, we suggest the prevention of C3a anaphylatoxin\u2010related reactions during immunoadsorption therapy.<\/jats:p>","DOI":"10.1002\/prca.200800232","type":"journal-article","created":{"date-parts":[[2009,7,10]],"date-time":"2009-07-10T03:22:18Z","timestamp":1247196138000},"page":"797-809","update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["MS characterization of apheresis samples from rheumatoid arthritis patients for the improvement of immunoadsorption therapy \u2013 a pilot study"],"prefix":"10.1002","volume":"3","author":[{"given":"Mike","family":"Kienbaum","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cornelia","family":"Koy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Helen V.","family":"Montgomery","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Susanne","family":"Drynda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter","family":"Lorenz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Harald","family":"Illges","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Koichi","family":"Tanaka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joern","family":"Kekow","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Reinhard","family":"Guthke","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hans\u2010Juergen","family":"Thiesen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael O.","family":"Glocker","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2009,7,9]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF00832003"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1136\/ard.52.Suppl_1.S39"},{"key":"e_1_2_6_4_2","volume-title":"54th Conference of the American Society for Mass Spectrometry and Allied Topics","author":"Deierling T.","year":"2006"},{"key":"e_1_2_6_5_2","first-page":"2131","article-title":"Effects of Prosorba\u00ae column apheresis in patients with chronic refractory rheumatoid arthritis","volume":"31","author":"Roth S.","year":"2004","journal-title":"J. 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