This paper investigates the bioseparation of binary
protein mixtures using polystyrene based anion exchange resin. Adsorption
experiments were conducted in batch mode using draft-tube internally recirculate
dair lift biocontactor in comparison with the conventional shake flask batch
adsorption equilibrium experiments. Binary protein mixtures contained bovine
serum albumin (BSA) and bovine haemoglobin (BHb) at different initial fractions.
Results from single solute adsorption experiments in biocontactor showed that
both proteins were equally adsorbed onto the resin with equilibrium reached in
an equal time period. This represents similar affinities towards the negatively
charged resin surface, although BSA was expected to adsorb through specific
forces. Adsorption results showed that BSA has hindered the BHb adsorption in
the biocontactor, although adsorption of both proteins was equally hindered in
the shake flasks adsorption experiments. Moreover, adsorption of BHb was
inhibited up to 29% in the presence of BSA compared to the adsorption of BHb
from a solution containing single solute of BHb at the same initial
concentration. Similarly, the presence of BHb has hindered the adsorption of
BSA by 59%. Adsorptions of both BSA and BHb from binary solution when each
formed 75% initial fraction while the other protein formed the remaining 25%
were relatively low with equilibrium reached in shorter time. Moreover,
considerable amount of proteins remained in the solution, which demonstrates
that multilayer adsorption most likely didn’t occur at the relatively small
protein concentrations used in the present study. In general, the higher
adsorption of BHb can also be related to the compressibility of its molecules
which allowed for higher adsorption capacity. The homogeneous and lower shear
environment in the airlift biocontactor compared to the other conventional
batch adsorption in shake flask reduced the compressibility of BHb that caused
higher BSA adsorption from binary solutions of BSA and BHb, which allowed for
better bioseparation of both proteins.
Cite this paper
Dahman, Y. and Jayasuriya, K. (2013) Preliminary study of binary protein adsorption system and potential bioseparation under homogeneous field of shear in airlift biocontactor. Advances in Bioscience and Biotechnology, 4, 710-718. doi: 10.4236/abb.2013.46094.
 Bickerstaff, G.F. (1997) Immobilization of enzymes and cells. Human Press, Totowa.
 Rapoza, R. and Horbett, T. (1990) The effects of concentration and adsorption time on the elutability of adsorbed proteins in surfactant solutions of varying structures and concentrations. Journal of Colloid and Interface Science, 136, 480-492. doi:10.1016/0021-9797(90)90395-5
 Van Holde, K.E., Johnson, W.C. and Ho, P.S. (1998) Principles of physical biochemistry. Prentice-Hall Inc., New Jersey.
 Denizli, A. and Piskin, E. (2001) Dye-legend affinity systems. Journal of Biochemical and Biophysical Methods, 49, 391-416. doi:10.1016/S0165-022X(01)00209-3
 Dahman, Y., Puskas, J., Margaritis, A. and Cunningham, M. (2004) Novel thymine-functionalized polystyrenes for applications in biotechnology. 2. Adsorption of Model Proteins. Biomacromolecules, 5, 1412-1421.
 Kondo, A. and Higashitani, K. (1992) Adsorption of model proteins with wide variation in molecular properties on colloid particles. Journal of Colloid and Interface Science, 150, 344-351.
 Shirahama, H., Susuki, K. and Suzawa, Y. (1989) Bovine haemoglobin adsorption onto polymer lattices. Journal of Colloid and Interface Science, 129, 483-490.
 Absolom, D., Zingg, W. and Neumann, A. (1997) Protein adsorption to polymer particles: Role of surface properties. Journal of Biomedical Materials Research, 21, 161171. doi:10.1002/jbm.820210202
 Yoon, J.Y., Park, H.Y., Kim, J.H. and Kim, W.S. (1998) Interpretation of protein adsorption phenomena onto functional microspheres. Journal of Colloids and Surface B, 12, 15-26. doi:10.1016/S0927-7765(98)00045-9
 Ghadge, R.S., Patwardhan, A.W., Sawant, S.B. and Joshi, J.B. (2005) Effect of flow pattern on cellulase deactivation in stirred tank bioreactors. Chemical Engineering Science, 60, 1067-1083. doi:10.1016/j.ces.2004.09.069
 Rathore, N. and Rajan, R.S. (2008) Current perspectives on stability of protein drug products during formulation, fill and finish operations. Biotechnology Progress, 24, 504514. doi:10.1021/bp070462h
 Jaspe, J. and Hagen, S.J. (2006) Do protein molecules unfold in a simple shear flow? Biophysical Journal, 91, 3415-3424. doi:10.1529/biophysj.106.089367
 Otero, C., Fernandez-Perez, M. and Perez-Gil, J. (2005) Effects of interactions with micellar interfaces on the activity and structure of different lipolytic isoenzymes from Candida rugosa. Enzyme and Microbial Technology, 37, 695-703. doi:10.1016/j.enzmictec.2005.03.023
 Wierenga, P.A., Egmond, M.R., Voragen, A. and de Jongh, H. (2006) The adsorption and unfolding kinetics determines the folding state of proteins at the air-water interface and thereby the equation of state. Journal of Colloid and Interface Science, 299, 850-857.
 Postel, C., Abillon, O. and Desbat, B. (2003) Structure and denaturation of adsorbed lysozyme at the air-water interface. Journal of Colloid and Interface Science, 266, 7481. doi:10.1016/S0021-9797(03)00571-X
 Garcia, M., Bonen, J., Ramirez-Vick, M. and Sadaka, A.V. (1999) Bioseparation process science. Blackwell, California.
 Ahuja, S. (2000) Handbook of bioseparations. Academic, California.
 Moo-Young, M. and Chisti, Y. (1989) On the calculation of shear rate and apparent viscosity in airlift and bubble column bioreactors. Biotechnology and Bioengineering, 34, 1391-1392. doi:10.1002/bit.260341107
 Chisti, Y. (1989) Airlift bioreactors. Elsevier, New York.
 Chisti, Y., Wenge, F. and Moo-Young, M. (1995) Relationship between riser and downcomer gas hold-up in internal-loop airlift reactors without gas-liquid separators. Chemical Engineering Journal, 57, 7-13.
 Peterson, E. and Margaritis, A. (2001) Hydrodynamic and mass transfer characteristics of three-phase gas-lift bioreactor systems. Critical Reviews in Biotechnology, 21, 233-294. doi:10.1080/07388550108984172
 Revilla, J., El-Aissari, A., Carriere, P. and Pichot, C. (1996) Adsorption of bovine serum albumin onto polystyrene latex particles bearing saccharidic moieties. Journal of Colloid and Interface Science, 180, 405-412.
 Causserand, C., Kara, Y. and Aimar, P.J. (2001) Protein fractionation using selective adsorption on clay surface before filtration. Journal of Membrane Science, 186, 165181. doi:10.1016/S0376-7388(01)00332-5
 Dickinson, E. (1999) Adsorbed protein layers at fluid interfaces: Interactions, structure and surface rheology. Journal of Colloids and Interfaces B: Biointerfaces, 15, 161-176.
 McMurry, J. (1999) Organic chemistry. 5th Edition, Brooks Cole, California.