Direct measurement of protein osmotic second virial cross coefficients by cross-interaction chromatography.
The importance of weak protein interactions, such as protein self-association, is widely recognized in a variety of biological and technological processes. Although protein self-association has been studied extensively, much less attention has been devoted to weak protein cross-association, mainly due to the difficulties in measuring weak interactions between different proteins in solution. Here a framework is presented for quantifying the osmotic second virial cross coefficient directly using a modified form of self-interaction chromatography called cross-interaction chromatography. A theoretical relationship is developed between the virial cross coefficient and the chromatographic retention using statistical mechanics. Measurements of bovine serum albumin (BSA)/lysozyme cross-association using cross-interaction chromatography agree well with the few osmometry measurements available in the literature. Lysozyme/alpha-chymotrypsinogen interactions were also measured over a wide range of solution conditions, and some counterintuitive trends were observed that may provide new insight into the molecular origins of weak protein interactions. The virial cross coefficients presented in this work may also provide insight into separation processes that are influenced by protein cross-interactions, such as crystallization, precipitation, and ultrafiltration.
Tessier
Peter M
PM
Center for Molecular and Engineering Thermodynamics, Department of Chemical Engineering, University of Delaware, Newark, Delaware 19716, USA.
Sandler
Stanley I
SI
Lenhoff
Abraham M
AM
D002845:Chromatography / Q000379:methods*; D002919:Chymotrypsinogen / Q000032:analysis / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000032:analysis / Q000737:chemistry; D008962:Models, Theoretical; D009113:Muramidase / Q000032:analysis / Q000737:chemistry; D009994:Osmolar Concentration; D011506:Proteins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000737:chemistry
D004800:Enzymes, Immobilized; D011506:Proteins; D012710:Serum Albumin, Bovine; D002919:Chymotrypsinogen; D009113:Muramidase