Biomedical subjects

R O Owen

Publications and source records attributed to R O Owen.

3 recordsLinked to original sources

PubMed1997

Preparation and use of ion-exchange chromatographic supports based on perfluoropolymers.

A poly(vinyl alcohol) (PVA) coated particulate perfluoropolymer (FEP) support has been functionalised with ion-exchange groups for use in ion-exchange chromatography of proteins. Anion-exchange (DEAE and Q) and cation-exchange (SP) groups were introduced to PVA-FEP which had previously been activated using cyanuric chloride. The equilibrium adsorption capacities of SP-PVA-FEP were 31.8 and 25.2 mg ml-1 for lysozyme and IgG respectively while for DEAE-PVA-FEP, the equilibrium adsorption capacities were 14.9 and 9.7 mg ml-1 for beta-lactoglobulin and HSA respectively. The equilibrium adsorption capacities for Q-PVA-FEP were determined to be 17.2 and 13.5 mg ml-1 for beta-lactoglobulin and HSA respectively. Experiments carried out to investigate the resolving power of materials showed that both SP and Q-PVA-FEP were able to resolve proteins with only small differences in their isoelectric points and that this resolution could be maintained at a flow-rate of 1500 cm h-1. SP-PVA-FEP was used to purify lysozyme from egg whites where a 50-fold purification, to homogeneity, was achieved in 98% yield. The anion exchanger, Q-PVA-FEP could be used to purify G6PDH from a clarified homogenate of bakers' yeast 14.3-fold in 81% yield.

Adsorption↗

PubMed1997

Direct purification of lysozyme using continuous counter-current expanded bed adsorption.

We describe the application of a novel technique for the continuous counter-current chromatography of proteins. The unit operation has shown potential in extracting targeted species from unclarified feedstocks, delivering clarified streams of purified product. The adsorbent used in this equipment consists of a perfluorocarbon matrix, coated with poly(vinyl alcohol), and derivatized with the triazine dye Procion Red HE-7B. Purification of lysozyme from egg-whites and enriched bovine milk could be carried out continuously. The former was extracted in 90.5% yield at a rate of 7400 U/min, achieving a purification factor of 19.4. Lysozyme from the enriched milk sample was extracted continuously at a rate of 41000 U/min, in 66.0% yield. The continuous products streams in both cases were fully clarified, thus enabling their direct application to a final polishing step, if desired.

Animals↗

PubMedDate not supplied

Application of perfluorocarbons in novel continuous counter-current protein chromatography.

In this work the development of a process capable of extracting proteins from particulate-containing solutions (such as fermentation broths) on a continuous basis, and in which absorbent and process streams are contacted counter-currently is described. The process consists of four stages, required for the loading, washing, elution and regeneration of the adsorbent. Because of its counter-current nature, it has improved performance over existing (though not yet commercialized) continuous processes, which have been based on CSTR-type contractors (e.g. PERCAS (McCreath et al., 1993). The improved efficiency has been shown by carrying out extraction of lysozyme from a single component feed stream. The adsorbent used in this work is a Procion Red HE-7B-derivatized perfluorocarbon support, which has shown particular suitability for continuous processes due to its inherently high mechanical strength and high density. Results illustrating yields obtained using this equipment are presented and discussed.

Adsorption↗