A novel approach to calculate protein adsorption isotherms by molecular dynamics simulations.

Protein adsorption plays a role in many fields, where in some it is desirable to maximize the amount adsorbed, in others it is important to avoid protein adsorption altogether. Therefore, theoretical methods are needed for a better understanding of the underlying processes and for the prediction of adsorption quantities. In this study, we present a proof-of-concept that the calculation of protein adsorption isotherms by molecular dynamics (MD) simulations is possible using the steric mass action (SMA) theory. Here we are investigating the adsorption of bovine/human serum albumin (BSA/HSA) and hemoglobin (bHb) on Q Sepharose FF. Protein adsorption isotherms were experimentally determined and modeled. Free energy profiles of protein adsorption were calculated by MD simulations to determine the Henry isotherms as a first step. Although each simulation contained only one protein, notably the calculated isotherms are in reasonably good agreement with the experimental isotherms. Hence, we could show that MD data can lead to protein adsorption data in good agreement with experimental data. The results were critically discussed and requirements for future applications are identified.

Expanding the elution by characteristic point method to columns with a finite number of theoretical plates.

The elution by characteristic point (ECP) method provides a rapid approach to determine whole isotherm data with small material usage. It is especially desired wherever the adsorbent or the adsorbate is expensive, toxic or only available in small amounts. However, the ECP method is limited to adsorbents that are well optimized for chromatographic use and therefore provide a high number of theoretical plates when packed into columns (2000 or more for Langmuir type isotherms are suggested). Here we present a novel approach that uses a new profile correction to apply the ECP method to poorly optimized adsorbents with less than 200 theoretical plates. Non-ideality effects are determined using a dead volume marker injection and the resulting marker profile is used to compensate the named effects considering their dependency from the actual concentration instead of assuming rectangular profiles. Experimental and literature data are used to compare the new ECP approach with batch method results.