Co-refining of fast pyrolysis bio-oil together with fossil oil in existing refinery infrastructure is an attractive and cost-efficient route to conversion of lignocellulosic biomass to transportation fuel. However, due to large differences in properties between the two oils, special notice is needed to reduce process-related issues. Here, fast pyrolysis bio-oil produced from lignocellulosic biomass was co-refined with vacuum gas oil at a 20:80 weight ratio in continuous operation in a pilot-scale slurry hydrocracker in order to investigate the impact of process parameters on product quality and process performance. Mass balances together with product characterization were used to investigate product yields, product composition, and hydrodeoxygenation. Best conversion and hydrodeoxygenation of the fast pyrolysis bio-oil was achieved using an unsupported catalyst loading of 900 ppm Mo with either a low temperature (410 °C) and long residence time (2 h) or higher temperature (435 °C) and shorter residence time (1 h). These settings resulted in about 94% hydrodeoxygenation but also led to highest yield of biogenic carbon to gas phase (40-43 wt %) and lowest yield of biogenic carbon to oil fractions (53-56 wt %) as well as the water fraction (3-5 wt %). Successfully, coke yield remained low at around 0.07-0.10 wt % for all performed runs, which was comparable to the insoluble particle content in the feed due to the presence of particles in the untreated fast pyrolysis bio-oil. Co-processing pyrolysis oil with fossil oil in a slurry hydrocracker seems to be a robust process with regard to coke formation, which should lead to reduced plugging issues compared to fixed bed hydrotreaters. Although this study gives a brief understanding of the effect of process parameters on the processing of fast pyrolysis bio-oil, further research is required to find optimal process parameters and to fully comprehend the possibilities and limitations for production of transportation fuels from fast pyrolysis bio-oil using this technology.