There is an incomplete understanding regarding the structural interactions of proteins with chromatographic resins used for protein separation and purification in the biotechnological industry, as the nanoscale architecture of chromatographic media has proven to be challenging to characterize using conventional techniques. We have used small-angle neutron scattering (SANS) as a novel application to measure (1) the structure of chromatographic media and (2) protein adsorption in these chromatographic media <i>in situ</i> with protein-scale resolution. The SANS spectra reveal how the architecture of the chromatographic media influences protein distribution upon adsorption, and, simultaneously, how protein adsorption can affect the architecture of the chromatographic media. In addition, protein-protein interactions of adsorbed protein at high loading levels show similarities to those of aggregated protein in solution. Based on these results, we have developed an analytical framework that can be used to study protein adsorption on chromatographic materials. These observations elucidate the structure and nature of protein partitioning in chromatographic materials to fundamentally understand their performance and aid in the design of novel, superior materials.

Influence of the architecture of chromatographic media on protein adsorption

253rd National Meeting (2017)

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