An integrated process for biomolecule isolation and purification.
Biomolecule isolation and purification from a fermentation broth usually involve centrifugation, filtration, adsorption, and chromatography steps. Each step contributes to the product cost and product loss. In this research, a cyclic process integrating commercially available ultrafiltration membranes and chromatographic resin beads was developed to achieve the same goal in one device. The device consisted of ion exchange beads on the shell side of a hollow fiber ultrafiltration module. Loading of proteins on the stationary phase on the shell side was carried out for a period of 5-20 min from the permeate on the shell side produced from tube-side feed in ultrafiltration. The eluent was then introduced either from the shell-side inlet or tube-side inlet; the chromatographic fractions were collected from the shell-side outlet. The column was regenerated/washed next to start a new cycle. Systems studied in this cyclic process include the following binary mixtures: myoglobin and beta-lactoglobulin; hemoglobin and bovine serum albumin; and myoglobin and alpha-lactalbumin. Excellent resolutions of the proteins were obtained. A yeast-based cellular suspension containing a mixture of myoglobin and alpha-lactalbumin was also applied to this device. The target proteins were recovered and purified successfully. The cyclic process-based device integrates clarification, concentration, and chromatographic purification of biomolecules and is suitable for both extracellular and intracellular products.
Dai
X P
XP
Center for Membrane Technologies, Department of Chemical Engineering, New Jersey Institute of Technology, Newark, New Jersey 07102, USA.
Luo
R G
RG
Sirkar
K K
KK
D000818:Animals; D001709:Biotechnology / Q000379:methods; D002417:Cattle; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods; D005285:Fermentation; D007768:Lactalbumin / Q000302:isolation & purification; D007782:Lactoglobulins / Q000302:isolation & purification; D009211:Myoglobin / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*; D012710:Serum Albumin, Bovine / Q000302:isolation & purification
D007782:Lactoglobulins; D009211:Myoglobin; D011506:Proteins; D012710:Serum Albumin, Bovine; D007768:Lactalbumin