Possibilities and Limitations of Different Separation Techniques for the Analysis of the Protein Corona.

One of the biggest challenges in the field of nanomedicine is the adsorption of biomolecules on the nanomaterial upon contact with a biological medium. The interactions of the so formed protein corona are essential for their behavior in a biological system. Thus, it is now commonly accepted that understanding the formation and consequently understanding the influence of the protein corona on the biological response is crucial. However, the outcome of the protein corona characterization cannot easily be compared between different studies and techniques, since many different sample preparation procedures exist that are suitable for different materials or methods. Depending on the applied procedure, the nanomaterial-protein-system will be altered in a certain way, so that it is necessary to consider the individual influence on the protein corona. Therefore, this review aims to give an overview on applied sample preparation methods for the analysis of the protein corona and to evaluate their influence on the outcome of the results especially with regard to the introduced terms "soft" and "hard protein corona". Special focus will be put on the comparison of the most commonly used techniques such as centrifugation, magnetic and chromatographic separation.

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