Correlation of diafiltration sieving behavior of lysozyme-BSA mixtures with osmotic second virial cross-coefficients.
The role of protein-protein interactions in membrane separations of protein mixtures remains incompletely understood, largely due to the difficulty of characterizing protein self- and, especially, cross-association. Recently, a novel technique, cross-interaction chromatography, has been developed to measure weak protein cross-association in terms of the osmotic second virial cross-coefficient. In this work the relationship between protein cross-association and the sieving behavior of lysozyme in the presence of BSA has been investigated. Sieving coefficients were measured using a stirred diafiltration cell over a range of pH and ionic strength, and a striking correlation between the lysozyme sieving and second virial cross-coefficients for BSA/lysozyme mixtures has been found: when the protein cross-interactions are most attractive (negative second virial cross-coefficient), the lysozyme sieving coefficients are lowest, and vice versa. The correlation between the sieving and second virial cross-coefficients may be due to the physically similar environments in the chromatography and filtration experiments since one protein is passed through a concentrated region of the second protein either immobilized on the column or accumulated at the membrane surface, and the migration rate of the mobile protein in both cases is influenced by protein cross-association. This study represents the first time that molecular interactions in binary mixtures have been related directly to filtration behavior, and may provide a useful approach to optimize the separation of other binary protein mixtures.
Center for Molecular and Engineering Thermodynamics, Department of Chemical Engineering University of Delaware, Newark, Delaware 19716, USA.
Peter M
PM
Tessier
Vincent J
VJ
Verruto
Stanley I
SI
Sandler
Abraham M
AM
Lenhoff
D045424:Complex Mixtures / Q000737:chemistry / Q000302:isolation & purification; D003956:Dialysis / Q000379:methods*; D044085:Microfluidics / Q000379:methods*; D008851:Micropore Filters; D046912:Multiprotein Complexes / Q000737:chemistry / Q000302:isolation & purification*; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification*; D009997:Osmotic Pressure; D016062:Porosity; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification*; D013223:Statistics as Topic; D014462:Ultrafiltration / Q000379:methods*
D045424:Complex Mixtures; D046912:Multiprotein Complexes; D012710:Serum Albumin, Bovine; D009113:Muramidase