TY - JOUR
T1 - Quantitative analysis of protein recovery from dilute, large volume samples by tangential flow ultrafiltration
AU - Powell, Matthew J.
AU - Timperman, Aaron T.
N1 - Funding Information:
We would like to thank the Flynn Research group at the Mary Babb Randolph Cancer Center for their generous donation of the E. coli cultures and hot room assistance. Also, we would like to thank James Lenke for his cooperative efforts in design and construction of the ultrafiltration system. The authors gratefully acknowledge the support of this research by the National Science Foundation EPSCoR Office, through a Research Infrastructure Improvement Award, RII-0314742.
Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 2005/4/15
Y1 - 2005/4/15
N2 - Important processes and parameters that affect the recovery yield using tangential flow ultrafiltration for the concentration and purification of proteins from dilute, large volume samples are investigated. Proteins from a radiolabeled E. coli cell lysate are concentrated from 60 and 20 L samples of artificial seawater. First, the improvement in the recovery and carryover with use of surface blocking agents is measured. The surface passivation resulted in a measurable decrease in adsorptive losses with minimal carryover. Additionally, the surface blocking agent pre-treatment resulted in more reproducible and reliable recovery yields (∼55-60%) and enhanced initial relative recovery by 46% for the initial 20 L processed. Second, the effects of the total protein sample concentration on the recovery yield are investigated. At low protein concentration (1.0 μg protein/L) the percent recovery yield was reduced, while at concentrations above 2.0 μg protein/L the percent recovery yield was nearly constant. Third, the components of the system were quantitatively analyzed to identify the relative contribution of the system components to adsorptive loss, and no single component was found to account for a large percent of the adsorptive loss. These results provide quantitative values for the recovery yields and critical information for improving protein recovery.
AB - Important processes and parameters that affect the recovery yield using tangential flow ultrafiltration for the concentration and purification of proteins from dilute, large volume samples are investigated. Proteins from a radiolabeled E. coli cell lysate are concentrated from 60 and 20 L samples of artificial seawater. First, the improvement in the recovery and carryover with use of surface blocking agents is measured. The surface passivation resulted in a measurable decrease in adsorptive losses with minimal carryover. Additionally, the surface blocking agent pre-treatment resulted in more reproducible and reliable recovery yields (∼55-60%) and enhanced initial relative recovery by 46% for the initial 20 L processed. Second, the effects of the total protein sample concentration on the recovery yield are investigated. At low protein concentration (1.0 μg protein/L) the percent recovery yield was reduced, while at concentrations above 2.0 μg protein/L the percent recovery yield was nearly constant. Third, the components of the system were quantitatively analyzed to identify the relative contribution of the system components to adsorptive loss, and no single component was found to account for a large percent of the adsorptive loss. These results provide quantitative values for the recovery yields and critical information for improving protein recovery.
KW - Dissolved proteins
KW - Membranes
KW - Radioactive labeling
KW - Surface blocking agents
KW - Ultrafiltration
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U2 - 10.1016/j.memsci.2004.12.024
DO - 10.1016/j.memsci.2004.12.024
M3 - Article
AN - SCOPUS:17144376762
VL - 252
SP - 227
EP - 236
JO - Jornal of Membrane Science
JF - Jornal of Membrane Science
SN - 0376-7388
IS - 1-2
ER -