Premium
Proteomics of methylene blue photo‐treated plasma before and after removal of the dye by an absorbent filter
Author(s) -
Crettaz David,
Sensebe Luc,
Vu DinhHao,
Schneider Philippe,
Depasse François,
Bienvenut WillyVincent,
Quadroni Manfredo,
Tissot JeanDaniel
Publication year - 2004
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200300667
Subject(s) - fibrinogen , transthyretin , chemistry , methylene blue , fibrin , chromatography , blood proteins , polymerization , apolipoprotein b , biochemistry , medicine , immunology , organic chemistry , cholesterol , polymer , catalysis , photocatalysis
Methylene blue (MB) and light are used for virus inactivation of plasma for transfusion. However, the presence of MB has been the subject of concern, and efforts have been made to efficiently remove the dye after photo‐treatment. For this study, plasma was collected by apheresis from 10 donors (group A), then treated using the MacoPharma THERAFLEX procedure (MB; 1 μ M , and light exposure; 180 J/cm 2 ) (group B), and finally filtered in order to remove the dye (group C). Proteins were analyzed by two‐dimensional electrophoresis, and peptides showing modifications were characterized by mass spectrometry. Clottable and antigenic fibrinogen levels, as well as fibrin polymerization time were measured. Analyses of the gels focused on a region corresponding to p I between 4.5 and 6.5, and M r from 7000 to 58 000. In this area, 387 ± 47 spots matched, and four of these spots presented significant modifications. They corresponded to changes of the γ‐chain of fibrinogen, of transthyretin, and of apolipoprotein A‐I, respectively. A decrease of clottable fibrinogen and a prolongation of fibrin polymerization time were observed in groups B and C. Removal of MB by filtration was not responsible for additional protein alterations. The effect of over‐treatment of plasma by very high concentrations of MB (50 μ M ) in association with prolonged light exposure (3 h) was also analyzed, and showed complex alterations of most of the plasma proteins, including fibrinogen γ‐chain, transthyretin, and apolipoprotein A‐I. Our data indicates that MB treatment at high concentration and prolonged illumination severely injure plasma proteins. By contrast, at the MB concentration used to inactivate viruses, damages are apparently very restricted.