<br/>The ability to characterize nanoparticles by asymmetric flow field-flow fractionation (AF4) has impacted a broad range of fields from biotechnology to nanotechnology. However, the availability of sufficiently large amounts of purified analytes is critical for conducting investigative studies concerning their fundamental properties and function. Precipitation and centrifugation have been used to isolate nanoparticles in quantities suitable for additional testing. However, precipitation may cause irreversible aggregation and sample collection after centrifugation is tedious. The development of a semi-preparative asymmetric flow field-flow fractionation (sP-AF4) method has the potential to overcome these limitations.<br/>The scale-up from an analytical to a semi-preparative channel means larger sample amounts can be introduced into the channel. However, increased channel size and sample amounts can impact separation resolution, destabilize channel flow rates, and increase potential membrane contamination. Separation performance metrics such as resolution and reproducibility are compared for analytical and sP-AF4 channels. Results showed that resolution is maintained in sP-AF4. Additionally, destabilization of flow rates and additional membrane contamination is not observed in the sP-AF4 channel.<br/>Purification of biological samples often results in low concentrations of the desired sample component making further analyses and studies difficult. A cancer-derived exosomes sample was fractionated using the sP-AF4 channel and SDS-PAGE of the collected fractions showed an abundance of exosome associated proteins. Fractionation of exosomes also revealed a distinct subpopulation of larger particles which may suggest the presence of non-exosome contaminants or exosome aggregates. The sP-AF4 channel was also used to produce more narrowly dispersed nanoparticles for subsequent investigations of enhanced nanoscale properties.

Semipreparative asymmetric flow field-flow fractionation for nanoparticle characterization

249th National Meeting (2015)

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