By UGA Biopharma…
UGA Biopharma patents superior method for EPO purification
Hennigsdorf, Germany: UGA Biopharma GmbH, contract developer of cell lines expressing biologics and biosimilars, has developed a newly patented method for the purification of the evolutionarily conserved hormone Erythropoietin (EPO) and its derivatives.
The innovative method and installation for purifying EPO and/or EPO derivatives was developed by two UGA employees, Wiebke Hannemann and Jenny Hessler. It makes the EPO purification process easier, more cost effective and environmentally friendly.
The patent for this method has been lodged with the European Patent Office and published as EP3613486A1 (See Resources).
Abstract and Summary:
The method and system described in the patent can be summarized as follows:
- The process uses a solid phase which has bound a 1-amino-2-sulfo-anthraquinone dye for immobilizing EPO and/or an EPO derivative. This measure achieves high purification of EPO and/or an EPO derivative to a desired purity while eliminating need for customary size exclusion chromatography, reversed-phase chromatography and/or addition of additives to the purification buffer. Thus, EPO and/or EPO derivative can be purified to high purity in a faster, simpler, more efficient, more cost-effective, and continuous manner. The patented method also lends itself to automation.
- For the purification of EPO and/or its derivatives, such as darbepoetin α, prior cleaning processes had at least four cleaning steps. These previously described methods used size exclusion chromatography (SEC) and/or reverse phase chromatography (RP), or buffers containing L-arginine to improve cleaning results.
- A disadvantage of SEC/RP is that they are unsuitable for fast, efficient and cost-effective cleaning of EPO/EPO derivatives on a large scale, since they have only very low loading capacities and long running times. RP has the additional drawback of regularly requiring organic solvents to be added to the running buffer, which is cost-intensive and environmentally harmful on an industrial scale and also represents a safety hazard (risk of explosion).
- The object of the newly invented method was to enable faster, simpler, more efficient and less expensive purification of EPO/EPO derivatives that could run continuously.
The patented purification comprises the following steps:
- contacting a liquid containing EPO and/or EPO derivative with a solid phase binding a 1-amino-2-sulfo-anthraquinone dye via non-covalent interactions
- cleaning the solid phase with a washing liquid
- eluting EPO/EPO derivative from the solid phase with an elution liquid
- reduction of the salt content of the elution liquid
- contacting of the reduced salt elution liquid with a solid hydroxyapatite material and collection of the run-through
- contacting of the (optionally pH-corrected) run-through with solid anion exchange material, whereby the EPO derivative binds to the solid anion exchange material
- washing of the solid anion exchange material with a washing liquid; and
- elution of the EPO and / or EPO derivative from the solid anion exchange material with an elution liquid.
- The UGA researchers found that using a solid phase, such as agarose, which has covalent bound a 1-amino-2-sulfo-anthraquinone dye, such as Cibacron blue from Sigma Aldrich, facilitates a strong immobilization of EPO/EPO derivatives, such as darbepoetin α, which consequently enables a complete removal of impurities from the liquid containing the EPO or EPO derivative. Therefore, use of SEC, RP or buffer additives are no longer necessary.
All together this means, the characteristic improvement of the method is therefore that EPO and / or an EPO derivative can be purified to a desired degree in a faster, simpler, more efficient and more cost-effective manner and the cleaning method can be operated continuously, which means an additional time and efficiency advantage.
About UGA Biopharma
UGA Biopharma GmbH is an experienced contract developer of biologics and biosimilars with the capabilities to handle all necessary steps from cell line development and bioprocess development to the development of purification strategies and appropriate analytics.
The company supplies its customers in Germany and abroad from its headquarter in Hennigsdorf, near Berlin. Several clients have used UGA products in clinical trials or in products that have received market authorization.
UGA Biopharma GmbH CRO activities are centered on cell line development for biologics and biosimilars, specializing in the development of stable expressing CHO cell lines and cell media for the production of biologics and biosimilars for biopharmaceutical companies. Typical clients are biopharma manufacturers, research institutions (like universities and research institutes) as well as start-ups and other CROs that need to outsource CLD, upstream, and/or downstream processing.
UGA’s mission is to fulfill customer requests, recognizing industry needs for high titer, high quality and fast delivery timelines, along with robust CHO cell lines. To this end, UGA can deliver cell lines, reaching titers from 4 to 7 g/L, within four months of receiving the originator sequence.
UGA has also developed its proprietary First CHOice® cell culture media and feeds platform to boost the productivity of cell lines. Furthermore, UGA Biopharma GmbH offers ready-to-use biosimilar cell lines, which allow clients interested in biosimilar development fast track access to a highly biosimilar cell line.
Further information at: www.ugabiopharma.com
Click on EPO purification patent description to see full details.