Membrane affinity chromatography is one of the processes that are considered as a possible alternative to bead based chromatography. Membrane chromatography is not limited by diffusion as the majority of chromatographic resins and therefore it is particularly suited for purifying large bio-molecules such as high molecular mass proteins and viruses. Nevertheless, in order to promote its applications in up-scaled plants, it is imperative to develop a reliable simulation tool able to describe the process performance in a predictive way. In this work we present a mathematical model that can describe a complete affinity cycle and its chromatographic steps: adsorption, washing and elution. The mathematical formalization takes into account all the relevant mass transport and kinetics phenomena involved in the membrane affinity chromatography process, namely axial convection, longitudinal dispersion in the micro-porous matrix and affinity binding with the specific adsorption site. Further, extra-column effects in terms of mixing volumes and delay time are included in the model. The few relevant fitting parameters were derived from a calibration with an extensive set of experimental affinity cycles performed with pure IgG solutions. The affinity cycles have been carried out using different innovative affinity membranes tested under a broad spectrum of operating conditions. Model validation is then achieved by comparing simulation results with experimental data which have been obtained for the purification of immunoglobulin G from a complex feed as a cell culture supernatant. Model simulations are in good agreement with the experimental affinity cycles both in the case of pure IgG solutions and for the cell culture supernatant considered, demonstrating the accuracy of the model to describe the transport phenomena in the column and the adsorption binding mechanism.

A mathematical model for the simulation of affinity membrane chromatography process

DIMARTINO, SIMONE;BOI, CRISTIANA;SARTI, GIULIO CESARE
2009

Abstract

Membrane affinity chromatography is one of the processes that are considered as a possible alternative to bead based chromatography. Membrane chromatography is not limited by diffusion as the majority of chromatographic resins and therefore it is particularly suited for purifying large bio-molecules such as high molecular mass proteins and viruses. Nevertheless, in order to promote its applications in up-scaled plants, it is imperative to develop a reliable simulation tool able to describe the process performance in a predictive way. In this work we present a mathematical model that can describe a complete affinity cycle and its chromatographic steps: adsorption, washing and elution. The mathematical formalization takes into account all the relevant mass transport and kinetics phenomena involved in the membrane affinity chromatography process, namely axial convection, longitudinal dispersion in the micro-porous matrix and affinity binding with the specific adsorption site. Further, extra-column effects in terms of mixing volumes and delay time are included in the model. The few relevant fitting parameters were derived from a calibration with an extensive set of experimental affinity cycles performed with pure IgG solutions. The affinity cycles have been carried out using different innovative affinity membranes tested under a broad spectrum of operating conditions. Model validation is then achieved by comparing simulation results with experimental data which have been obtained for the purification of immunoglobulin G from a complex feed as a cell culture supernatant. Model simulations are in good agreement with the experimental affinity cycles both in the case of pure IgG solutions and for the cell culture supernatant considered, demonstrating the accuracy of the model to describe the transport phenomena in the column and the adsorption binding mechanism.
2009 AIChE Annual Meeting and Fall Showcase Conference Proceedings on CD
1
1
S. Dimartino; C. Boi; G. Sarti
File in questo prodotto:
Eventuali allegati, non sono esposti

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11585/86839
 Attenzione

Attenzione! I dati visualizzati non sono stati sottoposti a validazione da parte dell'ateneo

Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus ND
  • ???jsp.display-item.citation.isi??? ND
social impact