Application of self-made asymmetric flow field flow fractionation instrument in protein separation / 中国药学杂志

ABSTRACT

OBJECTIVE: To design and construct aset of a symmetric flow field flow fractionation instrument which can be used to separate biomacromolecules, and preliminarily evaluate the instrument performance to provide practical experience for its application to real samples. METHODS: The apparatus was made up of separator and tempered glass. Polycarbonate membrane was used as the accumulated wall of the channel. In order to solve the problem of focusing samples, a novel focus position was developed. The performance of the asymmetric flow field flow fractionation device was preliminarily studied by injecting samples consisting of standard bovine serum albumin and alcohol dehydrogenase. The impacts of three factors at six levels on separation (resolution R) were investigated, including injection/focus time, cross-in flow and cross-out flow. RESULTS: Along with the decrease of focus time and increase of cross-in flow velocity, the resolution increased obviously. CONCLUSION: The self-made asymmetric flow field flow fractionation instrument can successfully separate protein mixtures and it is expected to separate more complex samples.