Self-interaction chromatography

The value of the second virial coefficient A 2 can also be determined from the retention volume that is obtained from self-interaction chromatography (Aliamed et al., 2005 Winzor et al., 2007 Dumetz et al., 2008). The chromatographic retention factor k is calculated from the retention volume VT using the formula... 

Ahamed, T., Ottens, M., van Dedem, G.W.K., van der Widen, L.A.M. (2005). Design of self-interaction chromatography as an analytical tool for predicting protein phase behaviour. Journal of Chromatography A, 1089, 111-124... 

Tessier, P.M., Lenhoff, A.M., Sandler, S.I. (2002). Rapid measurement of protein osmotic second virial coefficients by self-interaction chromatography. Biophysical Journal, 82, 1620-1631. 

Winzor, D.J., Scott, D.J., Wills, P.R. (2007). A simpler analysis for the measurement of second virial coefficients by self-interaction chromatography. Analytical Biochemistry, 371,21-25. 

In all modes of interactive chromatography, self-association of peptides and proteins can result in significant peak shape distortions, and RPC and HlC are no exception. Such behavior, often described in terms of isodemic interaction theory, is frequently a result of a specific molecular property of the peptide or protein, i.e., the propensity to form amphipathic helices or fibril-like structures or to selfassociate, and can lead to multiple peak zones for an otherwise compositionally homogeneous sample. Participation of such self-association processes can be assessed [127,128,338] by on-line light scattering (LALLS) spectroscopic procedures through an examination of the peak shape response as a function of sample concentration under specified RPC or HlC conditions at a defined pH and temperature. 

Self-associative lateral interactions can only occur with the AB-type analytes, chromatographed in sufficiently mild chromatographic conditions. In planar chromatography, this type of lateral interaction was first demonstrated on monocarboxylic fatty acids and a,co-dicarboxylic acids, chromatographed on microcrystalhne cellulose with aid of decalin and 1,4-dioxane as monocomponent eluents, respectively [8,20,23]. 

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