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Excess adapters were removed using an ultrafiltration column (Millipore, Billerica, MA).
Briefly, 5-ml urine was spun at 12,000 × g using an ultrafiltration column at 4 °C until approximately 1/30 of initial volume remained.
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Fractions containing the protein of interest, as judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE), were pooled and concentrated to ∼10 mg/mL using an ultrafiltration spin column with a YM30 membrane (Vivascience).
Micelles can then be separated from the emulsion using an ultrafiltration (UF) membrane.
Fractions that were positive for agarase activity were pooled and concentrated using an ultrafiltration concentrator.
The peptides were further purified from protein hydrolysates through using an ultrafiltration membrane with MWCO of 2 kDa.
The proteins were concentrated using an ultrafiltration device (VIVASpin).
Viscous food samples such as dressings and gravies were analysed using an ultrafiltration protocol [ 25].
Sera were individually albumin depleted as above and concentrated using a Vivaspin 2 ultrafiltration column with a 10 kDa molecular weight cut-off (GE Healthcare, Buckinghamshire, UK) according to the manufacturer's recommendations.
Total eluates were concentrated using Nanosep Ultrafiltration Columns (QIAGEN) down to a final volume of 50 μl.
The column was pre-equilibrated with 50 mM Tris-Cl pH 8, 150 mM KCl, 5 mM 2-mercaptoethanol and protein was eluted in the same buffer and was concentrated using Amicon ultrafiltration columns (Millipore).
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