Efficient capture of monoclonal antibody from cell culture supernatant using protein A media contained in a cuboid packed-bed device.

A box-shaped or cuboid packed-bed device was used for monoclonal antibody (mAb) separation using protein A affinity chromatography. The separation efficiency of the device was compared with an equivalent column i.e. packed with same resin, and having identical bed height and bed volume. The protein A media packed cuboid device had a larger number of theoretical plates than its equivalent column, e.g. 8750/m as opposed to about 4700/m at a flow rate of 0.5 mL/min. In mAb purification experiments, the impurity flow-through and eluted mAb peaks were shaper with the cuboid device. This implied that the effective separation time and buffer consumption with this device was lower, the purified mAb pooled volume was smaller, and the mAb concentration in the pooled volume was greater. Equivalent separation efficiency could be obtained with the cuboid device using higher flow rates than that used with the column. For instance, elution peaks equivalent to those obtainable by the column could be obtained at a 5 times greater flow rate using the cuboid device. The results discussed in this paper clearly demonstrate the potential for improving the efficiency of protein A affinity chromatography based mAb purification by using a cuboid packed-bed device.

Feasibility study for high-resolution multi-component separation of protein mixture using a cation-exchange cuboid packed-bed device.

In recent papers we have discussed the optimization of design and operating conditions for cuboid packed-bed device for chromatographic separations. The efficiency metrics used in these studies included the number of theoretical plates per unit bed height as well as attributes of flow-through and eluted peaks. These studies were carried out using equivalent columns as benchmarks. The cuboid packed-bed devices consistently outperformed the columns in terms of the above metrics. The current study examines how well, or indeed if at all these superior efficiency metrics translate to superiority in multi-component protein separation. Cation exchange resin was examined in the current study using appropriate multi-component model protein system which was chosen with close isoelectric points to make the separation challenging. Effects of operating and experimental parameters such as flow rate, loop size and linear gradient length on separation performance were systematically investigated. Separation metrics examined included peak width, tailing factor, asymmetry factor and resolution of separated protein peaks. The results obtained showed that the cation exchange cuboid packed-bed device significantly outperformed its equivalent commercial column (e.g., the number of theoretical plates per unit bed height was 8636/m for the cuboid packed-bed device as opposed to 1480/m for the column at a flow rate of 0.5 mL/min). The difference in efficiency was particularly high at lower flow rate and when shorter gradients were employed. The results suggest that the cuboid packed-bed devices could potentially have promising application in preparative separations such as biopharmaceutical purifications.