AlbuPure® Presentation

December 21, 2017

Hello, today I am going to talk to you about a custom designed affinity chromatography adsorbent for the biopharmaceutical industry. Prometic Bioseparations have over 25 years of experience and expertise in creating custom designed chromatography adsorbents with an extensive range of bioseparation products. Prometic provide cost effective solutions to enable the biomanufacturing industry and supply full technical support.

There are many protein-based therapeutics emerging in clinical trials for the treatment of a wide variety of complications, the issue however, is that once these biotherapeutics are delivered into the bloodstream, they will usually only last for hours, if not minutes, before it is removed and degraded. Biotherapeutic companies are therefore designing different technologies to ensure that their biomolecule is retained in the bloodstream to increase efficacy. Fusion technology is one such solution which is attracting a lot of attention in the biopharmaceutical industry as it can significantly increase the half-life of your biotherapeutic. As you can see in the example on this slide, extending the half-life of the biotherapeutic means that more stable blood levels can be attained, reducing side effects as well as the number of doses required, which is preferred by the patients and ultimately improves compliance.

As mentioned, one of the technologies available to increase the half-life of a biotherapeutic is Albumin fusion technology this is a platform technology which allows for any number of target biomolecules to obtain an increase half-life by engineering the target protein with human Albumin. On the slide, a schematic illustration of the model FC receptor mediated recycling pathway is presented and athelia cells lining the blood vessels allow for the uptake of Albumin by pinocytosis. The FC receptor binds Albumin in acidified recycling compartments, the complex is then recycled to the cell surface which results in exocytosis where the neutral pH of the bloodstream leads to the release of Albumin and its fused target biomolecule back into the bloodstream. Proteins that do not bind to the receptor will be sorted to lysosomes for degradation. This recycling of Albumin allows for Albumin fusion proteins to remain in the bloodstream increasing the half-life of the fusion partner.

The advantage Albumin fusion technology as a platform for increasing the efficacy of protein therapeutics also allows for a platform designed purification process. Prometic, in collaboration with Novozymes (now Albumedix) designed an adsorbent through our custom adsorbent program. This program includes an initial ligand discovery phase to identify suitable candidates, the next steps include adsorbent development, identifying the most suitable chemical attributes including ligand density, spacer arms and chemistry methodology. In parallel the adsorbent functionality is optimised in the chromatography development phase.

Finally, the optimised adsorbent is tech transferred to our Isle of Man manufacturing facility to complete the process validation. A full regulatory support package is created to accompany the final manufactured product.

The end goal was the manufacture of a novel synthetic adsorbent for the purification of Albumin fusion proteins, AlbuPure®. AlbuPure® is an affinity adsorbent which is highly stable, selective, robust and non-toxic, which is comprised of a novel colourless synthetic triazine ligand derived using PBL’s mimetic ligand technology and is coupled to our proprietary PuraBead® 6HF base matrix.

The following slides is a case study for the purification of interleukin 1 receptor antagonist Albumin fusion protein. Interleukin 1 receptor antagonist is an important regulator of the inflammatory and overall immune system. The native protein has a circulatory half-life of approximately six hours, the protein is used as a potential biomarker of disease states. As well as a biotherapeutic for the treatment of inflammatory diseases such as rheumatoid arthritis.

AlbuPure® was used to purify interleukin 1 receptor antagonist Albumin fusion protein from a yeast derived cell culture supernatant. The supernatant was pH adjusted to pH 5.3 and 20mg of target material was loaded per ml of AlbuPure®. Several incremental pH washes were carried out to remove minor non-selectively bound impurities. Finally the target material was eluted in a neutral buffer containing sodium octanoate. As AlbuPure® is caustic stable, the adsorbent was cleaned using a half molar sodium hydroxide.

This slide shows the chromatogram of the purification step.

SDS-page analysis of the process samples shows the majority of the impurities don’t bind and are washed through the column in the flow-through and post load wash steps. Minor non-target impurities are washed off using incremental pH washes as seen in lanes 4 and 5. Finally, the target material is eluted with high concentration and high purity.

In summary, Prometic Bioseparations in collaboration with Novozymes (now Albumedix) successfully designed a custom adsorbent for the purification of Albumin fusion proteins. In one instance interleukin 1 receptor antagonist Albumin fusion protein was purified using AlbuPure® obtaining a high purity and recovery. With these Albumin fusion partners, therapeutic proteins have shown a five-fold increase in their half-life which reduces the administration of therapeutic from daily to weekly injections.

Albumin fusion technology is an established technology with at least once licensed drug on the market and many more coming through in late phase clinical trials. Prometic not only meets industry requirements, providing a cost-effective purification platform technology for Albumin fusion proteins with AlbuPure® but also supplies off-the-shelf conventional and affinity adsorbents as well as quick screens and custom development programs to meet the needs of the biopharmaceutical industry.

Thank you for your interest, and if you require any further information on our custom-designed adsorbent services, AlbuPure® or any of our other products, please contact us using the information provided.

 

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