Studies on improved integrated membrane-based chromatographic process for bioseparation

To improve protein separation and purification directly from a fermentation broth, a novel membrane filtration-cum-chromatography device configuration having a relatively impermeable coated zone near the hollow fiber module outlet has been developed. The integrated membrane filtration-cum-chromatography unit packed with chromatographic beads on the shell side of the hollow fiber unit enjoys the advantages of both membrane filtration and chromatography; it allows one to load the chromatographic media directly from the fermentation broth or lysate and separate the adsorbed proteins through the subsequent elution step in a cyclic process. Interfacial polymerization was carried out to coat the bottom section of the hollow fiber membrane while leaving the rest of the hollow fiber membrane unaffected. Myoglobin (Mb), bovine serum albumin (BSA) and [alpha]-lactalbumin ([alpha]-LA) were used as model proteins in binary mixtures. Separation behaviors of binary protein mixtures were studied in devices using either an ultrafiltration (UF) membrane or a microfiltration (MF) membrane. Experimental results show that the breakthrough time and the protein loading capacities were dramatically improved after coating in both UF and MF modules. For a synthetic yeast fermentation broth feed, the Mb and u-LA elution profiles for the four consecutive cyclic runs were almost superimposable. Due to the lower transmembrane flux in this device plus the periodical washing-elution during the chromatographic separation, fouling was not a problem as it is in conventional microfiltration. A mathematical model describing the hydrodynamic and protein loading behaviors of the integrated device using UF membrane with a coated zone was developed. The simulation results for the breakthrough agree well with the experimental breakthrough curves. The optimal length of the coated zone was obtained from the simulation. A theoretical analysis of the protein mass transfer was performed using a diffusion-convection model considering the feed-side concentration polarization and the permeate-side concentration gradient formed by the adsorption. The permeate-side adsorption can enhance the observed protein transmission through the membrane considerably at low permeate flux. But the enhancement effect can be neglected at higher permeate flux when convection dominates the total mass transfer process or the proteins are very highly rejected by the membrane.

Name thesis:degree_name

Doctor of Philosophy in Chemical Engineering - (Ph.D.)

Discipline thesis:degree_discipline

Chemical Engineering

Year

2004

Author dc:creator

• Xu, Yanke

Contributors dc:contributor

• Kamalesh K. Sirkar
• Gordon Lewandowski
• Dana E. Knox

• Bioseparation
• Protein purification
• Chromatography
• Membrane filtration
• Interfacial polymerization
• Chemical Engineering

Repository record dc:identifier

https://digitalcommons.njit.edu/dissertations/616

OAI identifier oai:identifier

oai:digitalcommons.njit.edu:dissertations-1671