TY - JOUR
T1 - Novel internally staged ultrafiltration for protein purification
AU - Feins, Meredith
AU - Sirkar, Kamalesh K.
N1 - Funding Information:
The authors wish to thank the following sources for financial support: (1) Membrane Separations and Biotechnology Program at NJIT; (2) Otto H. York Department of Chemical Engineering of NJIT; (3) Center for Membrane Technologies at NJIT.
PY - 2005/2/15
Y1 - 2005/2/15
N2 - A new ultrafiltration technique based on a multimembrane stack has been developed to fractionate proteins closer in molecular weight than conventionally possible. The technique is illustrated here by obtaining a pure protein product from a binary protein mixture. By employing membranes in series using the same membrane without any gaskets or spacers in-between, ultrafiltration is carried out to separate two proteins relatively close in molecular weight. Either flat YM100 regenerated cellulose membranes or Omega 100 K polyethersulfone membranes, of the same molecular weight cutoff (MWCO) 100,000, are stacked together in the desired number, and ultrafiltration takes place. The membrane rejection of a protein is amplified with each additional membrane, ultimately resulting in a completely rejected species. Complete purification of the more permeable protein may be achieved by operating under a physicochemical condition that is optimal for selective separation by a single membrane. The separation of hemoglobin (MW 64,677) and bovine serum albumin (BSA, MW 66,430) was studied under various operating conditions; the molecular weight ratio is 1.03. Complete rejection of bovine serum albumin was achieved using three Omega 100 K membranes one on top of the other. To achieve complete rejection in a multimembrane stack, the single membrane rejection must be considerable. Cleaning in situ was achieved with reproducible experimental results before and after on-line cleaning. The results clearly demonstrate that multimembrane stacks can be used for fractionation of proteins that are quite close in molecular weight.
AB - A new ultrafiltration technique based on a multimembrane stack has been developed to fractionate proteins closer in molecular weight than conventionally possible. The technique is illustrated here by obtaining a pure protein product from a binary protein mixture. By employing membranes in series using the same membrane without any gaskets or spacers in-between, ultrafiltration is carried out to separate two proteins relatively close in molecular weight. Either flat YM100 regenerated cellulose membranes or Omega 100 K polyethersulfone membranes, of the same molecular weight cutoff (MWCO) 100,000, are stacked together in the desired number, and ultrafiltration takes place. The membrane rejection of a protein is amplified with each additional membrane, ultimately resulting in a completely rejected species. Complete purification of the more permeable protein may be achieved by operating under a physicochemical condition that is optimal for selective separation by a single membrane. The separation of hemoglobin (MW 64,677) and bovine serum albumin (BSA, MW 66,430) was studied under various operating conditions; the molecular weight ratio is 1.03. Complete rejection of bovine serum albumin was achieved using three Omega 100 K membranes one on top of the other. To achieve complete rejection in a multimembrane stack, the single membrane rejection must be considerable. Cleaning in situ was achieved with reproducible experimental results before and after on-line cleaning. The results clearly demonstrate that multimembrane stacks can be used for fractionation of proteins that are quite close in molecular weight.
KW - Biotechnology
KW - Composite membranes
KW - Internally-staged flat membranes
KW - Protein purification
KW - Ultrafiltration
UR - http://www.scopus.com/inward/record.url?scp=12344296469&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=12344296469&partnerID=8YFLogxK
U2 - 10.1016/j.memsci.2004.09.035
DO - 10.1016/j.memsci.2004.09.035
M3 - Article
AN - SCOPUS:12344296469
SN - 0376-7388
VL - 248
SP - 137
EP - 148
JO - Journal of Membrane Science
JF - Journal of Membrane Science
IS - 1-2
ER -