Protein Purification and Production Services

The growing global need to produce large amounts of well-defined biological products, including recombinant proteins such as monoclonal antibodies, cytokines and subunit vaccines, as diagnostics and/or therapeutics for human use has necessitated the development of efficient large-scale purification processes to generate safe and reproducible materials at reasonable cost. The key to achieving efficient and robust purification procedures is selection of the most appropriate chromatographic resins for each separation step comprising a multidimensional process, the sequence of which is critical for optimisation of chromatographic performance in terms of simplicity, scalability and product yield.

The Monash site of the Victorian Node Recombinant Protein Feeder Facility houses state-of-the art equipment for automated high throughput screening to develop and optimise efficient protein purification schemes and complementary analytical/characterisation methods that result in:

• Final product in high purity
• Maximum yield
• Process economy

Upstream Processing:

• Fermentation
• Cell culture
• Harvesting
• Filtration and concentration

Downstream Processing:

• Synthesis of novel ligands for affinity chromatography
• High throughput ligand and resin screening
• Method development and optimisation
• Laboratory scale protein purification
• Pilot scale protein purification
• Process scale protein purification
• Ultrafiltration/Diafiltration
• Protein characterisation
• QC/QA

Purification of Recombinant Proteins

Chromatographic purification of proteins typically involves an initial capture step using a high-capacity resin for rapid processing of large volumes of starting material to concentrate the target protein, then one or more high-performance affinity-based chromatographic steps to resolve the target protein at reasonable to high levels of purity. A final polishing step yields the target protein at the highest possible purity.

High-throughput Resin Screening and Method Development

The selectivity of the resolving step(s) is determined by the specificity of interactions between the target molecule and various affinity resins. In recent years, new chromatographic media have been developed which possess good physical and chemical characteristics that permit (i) high flow-rates and (ii) good resolution that are readily adaptable across a broad range of process scales for the production of recombinant proteins. Considerable effort has also been focused on the development of novel small molecular weight ligands that have defined selectivity for specific classes of proteins, and which can be used in new affinity-based chromatography media.

The NCRIS Victorian Node Recombinant Protein Feeder Facility at Clayton is a national level capability that will exploit the use of classical chromatographic techniques for scale-up of protein purification coupled with expertise in the development of novel ligands for new modalities of affinity chromatography. Robotic, automated high throughput screening of immobilised ligands as affinity chromatographic adsorbents will provide rapid route scouting for method development, optimisation, process intensification and validation by implementing the concept of ‘scaling-down’ to effect efficient scale-up. This facility provides a unique combination of platform technologies and capabilities in a powerful combination for the development of more efficient, sustainable and cost-effective large-scale protein purification.

Users of this facility will benefit from the specialised services, expertise and state-of-the-art equipment and technologies that are provided in order to save time and costs when recombinant protein purification is required. Researchers at universities, other research institutions and SMEs will have subsidized access to services for upstream mammalian cell culture and microbial fermentation at CSIRO‘s Division of Molecular and Health Technologies and downstream protein purification at Monash University for various scales of production, ranging from small laboratory scale quantities through to larger process scale quantities depending upon specific requirements. Larger biotechnology companies and other industry groups will have access to these facilities at costs that reflect commercial rates.