Packing quality, protein binding capacity and separation efficiency of pre-packed columns ranging from 1 mL laboratory to 57 L industrial scale

摘要

Pre-packed chromatography columns are routinely used in downstream process development and scale-down studies. In recent years they have also been widely adopted for large scale, cGMP manufacturing of biopharmaceuticals. Despite columns being qualified at their point of manufacture before release for sale, the suitability of pre-packed chromatography columns for protein separations at different scales has not yet been demonstrated. In this study, we demonstrated that the performance results obtained with small scale columns (0.5 cm diameter x 5 cm length, 1 mL column volume) are scalable to production sized columns (60 cm diameter x 20 cm length, 57 L column volume). The columns were characterized with acetone and blue dextran pulses to determine the packing density and packed bed consistency. Chromatography performance was evaluated with breakthrough curves including capacity measurements and with separation of a ternary protein mixture (lysozyme, cytochrome C and RNase A) with a step gradient. The equilibrium binding capacity and dynamic binding capacity were equivalent for all columns. The step gradient separation of the ternary protein mixture displayed similar peak profiles when normalized in respect to column volume and the eluted protein pools had the same purities for all scales. Scalable performance of pre-packed columns is demonstrated but as with conventionally packed columns the influence of extra column volume and system configurations, especially buffer mixing, must be taken into account when comparing separations at different scales. (C) 2019 The Authors. Published by Elsevier B.V.