Field-flow fractionation (FFF) is an elution-based separation technique which is capable of the rapid and efficient separation of macromolecules, colloids and particles. Asymmetrical flow FFF (AF4) is one of the FFF sub-techniques that has been broadly used in characterization of complex biological and pharmaceutical products. The commercial AF4 system commonly employs flat channels with different dimensions and aspect ratios (10-28 cm long and 2-10 cm wide). The AF4 channel could be further downscaled to reduce channel volume (less dilution), sample consumption and operating cost. A miniaturized flat AF4 channel (microchannel) with a size smaller than a credit card was constructed and examined for characterization of biological samples spanning a wide range of molecular weight and diameter. The performance of the microchannel was tested using a mixture of plasma proteins. The resolution and reproducibility of the microchannel were found to be similar to those of a standard size channel. The microchannel was used to fractionate Human Serum Albumin (HSA) from different lipoprotein fractions. Baseline separation was also achieved between high-density and low density lipoproteins. An AF4 microchannel is an alternative to full-size channels which allows use of smaller sample mass, and reduces the amount of dilution in the channel, allowing more efficient detection.<br/>

Characterization of plasma proteins and lipoproteins using microchannel asymmetrical flow field-flow fractionation

254th National Meeting (2017)

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