Hollow fiber flow field-flow fractionation
Coriolis Pharma offers hollow fiber flow field-flow fractionation (HF5) contract laboratory services for aggregate content and molecular weight distribution as an orthogonal method to size exclusion chromatography or analytical ultracentrifugation for characterization of biopharmaceuticals.
Hollow fiber flow field-flow fractionation (HF5) is a recently introduced field-flow fractionation technique, offering certain advantages over the traditionally used AF4 method. Similar to AF4, separation is achieved by applying a cross-flow, but instead of using a channel a hollow fiber cartridge is used for the separation. Consequently, in HF5 a radial cross-flow towards the porous fiber wall is applied, whereas AF4 uses a transverse, unidirectional cross-flow towards a flat membrane. A wide size range from several nm to µm can be covered by HF5, which is a clear advantage compared to HP-SEC. At Coriolis, HF5 can be offered in full GMP compliance.
The primary benefit of HF5 separation is its high sensitivity, owing to the low channel volume (typically < 100 µl). Thus, the hollow fiber devices entail a low dead volume, minimal solvent and sample requirements and the separation is very fast. Thus, HF5 is especially useful for method development and early stage analysis. An established method can later on easily be transferred to AF4 channels, allowing a higher sample load.
Similar to AF4 and HP-SEC, HF5 can be combined with UV, fluorescence, RI and MALLS detection for a broad variety of biopharmaceutical applications. Applications are similar to AF4, e.g., during forced degradation studies (see relevant publication) and real-time stability studies (see relevant publication). Furthermore, as an orthogonal method to HP-SEC HF5 is of increasing importance for batch release with respect to the cross-validation of size-exclusion chromatography methods to characterize biopharmaceutical formulations.
At Coriolis, numerous AF4 and HF5 methods for peptides/proteins, virus-like particles, liposomes and polymers have been successfully developed.