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Plasminogen activator inhibitor-1 (PAI-1) is a member of the serine protease inhibitor (serpin) superfamily and is the principal inhibitor of the plasminogen activators tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA) in vivo. In healthy individuals, PAI-1 is found at low levels in the plasma, but is elevated significantly in a number of diseases, including atherosclerosis, deep vein thrombosis, and non-insulin dependent diabetes mellitus. Objective: Elucidation of the molecular interaction mechanism between PAI-1 and a PAI-1 inhibiting antibody fragment Fab-55F4C12. Methods: Fab-55F4C12 was generated by papain digestion of MA-55F4C12, followed by protein A and gelfiltration purification. The purified Fab-55F4C12 was concentrated to OD = 10. Different crystallisation screens were tried, and initial crystal clusters were obtained in condition 19 of Structure Screen 1 of Molecular Dimensions. Small needles (0.2 x 0.1 x 0.1 mm) were obtained after intensive optimisation of the crystallisation condition. The crystal structure of Fab-55F4C12 was determined by X-ray crystallography at cryogenic temperature. The data set was collected at DESY (Hamburg, Germany) to a resolution of 2.7 Å. Data processing was done using MOSFLM and SCALA. The space group was assigned to be P21212 with unit-cell parameters a = 52.04 Å, b = 98.66 Å, c = 191.68 Å, with two molecules in the asymmetric unit. The data set is 99.72 % complete. Initial phases were obtained with molecular replacement. The structure was refined using Coot and Refmac5. Crystallisation of the Fab-55F4C12 / PAI-1 complex has been unsuccessful so far. Therefore, the complex is being modelled through docking of the crystal structures of the two subunits of the complex using the rigid-body docking programs DOT and ZDOC. Potential interactions between the two subunits will be deduced from the models and compared with epitope information gathered from mutagenesis studies. Conclusions: Characterization of the complex of Fab-55F4C12 with PAI-1 may provide valuable information on the molecular interactions between the Fab-fragment and PAI-1, leading to a better understanding of the mechanism of inhibition. The elucidation of the binding site of inhibitory monoclonal antibodies may contribute to the rational design of PAI-1 modulating therapeutics. m06.p03