The primary aim of the study was to develop and validate an in-house upscale of Automatic Methane Potential Test System II for studying real-time inocula and real-scale substrates in batch, codigestion and enzyme enhanced hydrolysis experiments, in addition to semi-continuous operation of the developed equipment and experiments testing inoculum functional quality. The successful upscale to 5 L enabled comparison of different process configurations in shorter preparation times with acceptable accuracy and high-through put intended for industrial decision making. The adoption of the same scales, equipment and methodologies in batch and semi-continuous tests mirroring those at full scale biogas plants resulted in matching methane yields between the two laboratory tests and full-scale, confirming thus the increased decision making value of the approach for industrial operations. (C) 2013 Elsevier Ltd. All rights reserved.