Journal: Journal of thrombosis and haemostasis : JTH

Article Title: The lectin complement pathway serine proteases (MASPs) represent a possible crossroad between the coagulation and complement systems in thromboinflammation.

doi: 10.1111/jth.13208

Figure Lengend Snippet: Fig. 4. Panels A–E: binding of LP components from serially diluted lepirudin-PPP (starting with 3% dilution) to plasmin-generated fragments of fibrinogen and fibrin coating microtiter plates. Detection was carried out using specific biotinylated mAbs followed by streptavidin-HRP (n = 3). Panel A: negligible binding of MBL, Ficolin-1, Ficolin-2 and Ficolin-3 to fibrin fragment DD. Panel B: significant binding of MASP-1 and, to a lesser degree, MASP-2 to fibrin fragment DD. Insignificant binding of MASP-1 and MASP-2 to fibrin fragment E (panel C), fibrino- gen fragment D (panel D) and fibrinogen fragment E (panel E). Panels F–G: generation of MASP-1 and MASP-2/AT complexes by fibrin and fibrin fragment DD. Lepirudin-PPP (n = 3) was incubated with increasing concentrations of fibrin (0–12 lg mL1, panel F) or fibrin fragment DD (0–25 lg mL1, panel G). After incubation MASP-1 and MASP-2/AT/C1-INH complexes were quantified by sandwich ELISA. In PPP in contact with fibrin, MASP-1/AT and MASP-2/AT complexes were preferentially formed (panel F). Fibrin fragment DD induced similar genera- tion of both MASP-1, MASP-2/AT and MASP-1 and MASP-2/C1-INH, but the latter were formed at a much lower concentration of DD (panel G). LP, lectin pathway; PPP, platelet-poor plasma; MBL, mannose-binding lectin; AT, antithrombin.

Article Snippet: 4 The wells were then incubated with 60 lL of HRP-conjugated streptavidin (GE Healthcare) diluted 1 : 500.

Techniques: Binding Assay, Generated, Incubation, Sandwich ELISA, Concentration Assay, Clinical Proteomics