Enthalpy contributions to adsorption of highly charged lysozyme onto a cation-exchanger under linear and overloaded conditions

• FMC powerful technique to illustrate protein adsorption mechanisms in IEC.
• Direct correlation between microcalorimetry data and isotherm measurements.
• Under studied conditions adsorptive IEC process is enthalpy driven.
• Great contribution from observed entropy resulting from desolvation processes.
• Entropy contribution overcome by exothermic heat due to protein reorientation.

An investigation of the adsorption mechanism of lysozyme onto carboxymethyl cellulose (CMC) was conducted using flow calorimetry and adsorption isotherm measurements. This study was undertaken to provide additional insight into the underlying mechanisms involved in protein adsorption that traditional approaches such isotherm measurements or van’t Hoff analysis can’t always provide, particularly when protein adsorption occurs under overloaded conditions. Lysozyme and CMC were selected for this study because the characteristics of the protein and the adsorbent are well known, hence, allowing the focus of this work to be on the driving forces influencing adsorption. Calorimetry results have showed that lysozyme adsorption onto CMC produced both exothermic and endothermic heats of adsorption. More specifically flow calorimetry data coupled with peak deconvolution methods illustrated a series of chronological events that included dilution, primary protein adsorption, rearrangement of surface proteins and a secondary adsorption of lysozyme molecules. The observations and conclusions derived from the experimental work presented in our figures and tables were developed within the mechanistic framework proposed by Lin et al., J. Chromatogr. A. 912 (2001) 281.