The continual depletion of fossil fuel resources necessitates sustainable solutions to face the global demand for energy, fuels and chemicals. Pyrolysis followed by hydrotreatment offers an attractive and economical strategy for thermochemical conversion of biomass to fuels and chemicals. However, optimisation of the hydrotreatment process and catalyst properties is still necessary to promote it to an industrial scale. This study presents a comprehensive analysis of nickel-based catalysts for hydrotreatment of pyrolysis oil in mild and deep reaction conditions. In-depth catalyst characterisation showed the influence of pyrolysis oil on the catalyst activity. Finally, a systematic analysis about the influence of different parameters on the upgraded oil composition from intermediate pyrolysis oil is reported.