An annular-flow, hollow-fiber membrane chromatography device for fast, high-resolution protein separation at low pressure

High-resolution protein separation is currently carried out using columns packed with ultrafine particulate chromatographic media which can only be operated at very high pressures. This restricts the flow rate and thereby the speed of separation. A membrane with ultrafine pores shares the same mass transport advantages as a bed of ultrafine particles. However, application of membrane chromatography has largely been restricted to low-resolution separation applications, primarily due to limitations associated with poorly designed devices. In this paper, we describe a novel annular-flow hollow-fiber membrane chromatography (AHMC) device, suitable for fast, high-resolution protein separation. The AHMC device was designed primarily from a fluidics perspective to ensure a narrow solute residence time distribution, a low dead-volume, and minimal fluid back-mixing. Using this device, it was possible to separate a ternary protein mixture in less than a minute, at less than 0.3 MPa backpressure. The high-resolution separation capability of AHMC was further demonstrated through its use for the separation of bovine serum albumin (BSA) dimer from the monomeric form of the protein. This was achieved in less than 3 min which was considerably faster than other reported techniques for protein aggregates separation.