Continuous Radial Flow Chromatography of Proteins

Abstract

A continuous radial flow chromatography (CRFC) system consisting of an annular packed bed between two rotating concentric porous stainless steel cylinders was constructed and tested. The protein feed, wash buffer and elution buffer are applied simultaneously at different fixed angular positions at the periphery of the rotating bed and flow radially inwards. The target protein is bound in the feed zone and eluted in the elution zone while unbound protein flows through the feed zone. Continuity equations for fixed bed radial flow chromatography were extended to model the CRFC by including angular displacement terms, and solved using finite difference methods. Film diffusion at the resin particle boundary and pore diffusion within the resin particle was described by an overall mass transfer coefficient and a multicomponent Langmuir isotherm was used to describe protein absorption onto the resin particle. To verify the model, a solution containing 0.53 mg ml 21 lactoferrin and 1.6 mg ml 21 bovine serum albumin (BSA) was applied at 5 ml min 21 to a 220-ml CRFC system packed with DEAE Sepharose Fast Flow ion exchange resin, with abed speed of 0.02 rpm. The proteins were separated into lactoferrin of 68% purity (74% recovery) and BSA of 94% purity (85% recovery). Experimental data were used to find values for the parameters in the model proposed.

Study on the purification of polysaccharides from Noscoc flagelliforme with radial flow chromatography

“…It was proved to be a promising alternative to conventional axial flow chromatography (AFC) [5]. Its relatively large flow area and short flow path allow fast separations and low pressure drops [6,7] and it has been used for a wide range of separations [8][9][10][11][12][13]. An experimental case study of the comparison of RFC and AFC was carried out by Saxena and Weil [14] for the separation of ascites.…”

Integrated system for temperature-controlled fast protein liquid chromatography. II. Optimized adsorbents and ‘single column continuous operation’

“…Unless fully optimized, a common feature is inefficient use of the separation medium. The switch from batch to continuous operation promises several advantages, key of which is more efficient utilization of the bed [43]. Several continuous chromatography formats have been developed and applied for the separation of biomacromolecules thus far, including Continuous Annular Chromatography [44], Continuous Radial Flow Chromatography [43], Simulated Moving Bed in various guises [45][46][47][48][49], and Periodic Counter-current Chromatography [50].…”

Perspectives on integrated continuous bioprocessing—opportunities and challenges

“…These systems provide periodic (cyclical) operation, with the product concentration in the eluent stream varying with time during each cycle. Annular [28] and radial [29] chromatography can also be operated in continuous mode with true steady-state operation, although neither of these have been implemented for large-scale commercial protein purification.…”