Bioprocessing feature: Out of the vat

Bioprocessing and fermentation are essential parts of the biotechnology product development cycle. If a drug or therapeutic is produced by a living organism, whether it's bacteria, yeast, plant cells or mammalian cells, some form of large-scale fermentation or culture is required to produce the entity, and usually some form of downstream processing is necessary to extract, purify, concentrate and formulate the end product.

The need for bioprocessing is not restricted to companies preparing a product for pre-clinical or clinical trials. The development of bioprocesses often starts in a research environment, with the expression of a protein of interest in E. coli, or the development of a monoclonal antibody cell line. But once the scale of bioprocessing increases to a size where research lab protocols are no longer sufficient, many companies turn to specialised contract manufacturing facilities to assist them in developing their bioprocessing needs, and in many cases to manufacture Good Manufacturing Practice (GMP) compliant products.

Few Australian companies have their own bioprocessing development and manufacturing facilities, with the exception of CSL, which has extensive development and manufacturing facilities, and some of the companies with successful products, such as GroPep and BresaGen. Part of this is due to the cost of setting up such a facility, as well as the relative immaturity of the Australian biotech sector.

So when GroPep closed the former Biotech Australia contract manufacturing facility earlier this year, one of the few options for out-sourced bioprocessing in Australia was lost to the sector. There are only a handful of facilities in the country that can handle contract bioprocessing and manufacturing and these facilities have varying capacities and strengths. Among these are no less than three groups at CSIRO, including a tissue culture facility at CSIRO Health Sciences and Nutrition in Parkville (Victoria), bacterial, yeast and fungal fermentation and processing at CSIRO Molecular Sciences in Clayton (Victoria), and plant cell culture facilities under the wing of Food Science Australia in its North Ryde (NSW) and Werribee (Victoria) facilities.

Private sector contract manufacturers include Progen Industries with facilities for bacterial, yeast and fungal fermentation and some mammalian cell culture too, and BresaGen, which has an E. coli-based expression system and contract manufacturing facilities. Alpharma also has the capacity to do some bioprocessing on a contract basis. Straddling the public and private sectors is Acyte, a spin-off from the Bioengineering Centre at the University of NSW, offering mammalian cell culture expression systems and bioprocessing services. Surprisingly, the major users of most of these bioprocessing facilities come from overseas, mainly from the US and Europe.

Global shortage

"There is a global biomanufacturing shortage, especially for monoclonal antibodies, and it's only going to get worse," says Dr Michael Zachariou, who heads CSIRO Molecular Sciences' fermentation Facility. He notes that the lack of available facilities was a major talking point at the Bio 2002 conference in Toronto earlier this year. Zachariou believes that Australia could be in a good position to form a consortium of biomanufacturing and bioprocessing facilities to market to the international biotechnology and pharmaceutical industry, particularly given the cost-effectiveness of manufacturing bioproducts in Australia.

"Australia is too small to try to have too many of these industries running on its own. We should present to the international scene as a unified bioprocessing nation," he explains. In a step in that direction, Zachariou's group at CSIRO has allied itself with Progen to provide a wide range of fermentation and bioprocessing services, with the CSIRO group concentrating on earlier-stage and smaller-scale development and manufacturing, and Progen providing GMP-compliant pilot-scale facilities.

"Because we have the knowledge of the GMP environment and what it takes to develop GMP processes, we can develop a GMP process quite quickly and then transfer it to Progen for GMP production," Zachariou says. Progen is also able to provide any necessary quality assurance for the CSIRO group. Between the two facilities, the capacity for bacterial and yeast fermentation ranges from 2 litres to 700 litres. They also have extensive capabilities for developing downstream processing techniques, such as continuous flow homogenisers and centrifuges, tangential flow concentration and clarification systems, separative chromatography and analytical abilities.

Progen's facility is licensed by the Therapeutic Goods Administration (TGA) and the National Registration Authority (NRA -- which oversees veterinary product manufacture) as well as the US Food and Drug Administration (FDA). "As far as I know, we are the only active licensed biopharmaceutical manufacturing facility in Australia at the moment," says Progen's Operations supervisor Les Tillack. "I find it hard to understand why there aren't more facilities with the number of products being talked about."

Adelaide-based BresaGen also provides contract bioprocess development and manufacturing services, specialising in producing smaller proteins and peptides in E. coli. "We're pushing a new approach called protEcol Services. It's a new service that BresaGen's protein pharmaceuticals division offers and includes a process development service," explains R&D manager Dr Stan Bastiras. The company's facility is TGA and NRA registered and has a maximum capacity of up to 100 litres per fermentation run, which Bastiras says is sufficient for development activities and manufacture of pre-clinical and clinical material for use up to Phase II.

Next year BresaGen will open a new facility with a 500 litre fermentation capacity, which will meet FDA specifications for GMP compliance. According to Bastiras, this will allow BresaGen to go beyond the current capabilities and provide material for Phase III trials and beyond. Bastiras doesn't see the need in Australia for a consortium approach to contract process development and manufacturing, citing the difficulties in reconciling the needs of public and commercial sector interests. He believes that bioprocessing is a niche opportunity which runs mainly by word of mouth recommendations and repeat customers. "Word spreads and people come back to you," he says.

Mammalian culture is lacking

Among the facilities, mammalian cell culture seems to be the worst off for facilities in Australia, with little or no contract bioprocessing and manufacturing at the GMP-compliant end of the scale. "I think we're the only ones to do large scale mammalian cell culture," says Dr George Lovrecz, who leads the Fermentation Group at CSIRO Health Sciences and Nutrition. He says their facility houses spinner flasks, roller bottles and bioreactors, with a maximum capacity of 200 litres in a continuous perfusion mode, and can work with mammalian cells including hybridomas as well as insect baculovirus cultures.

But the facility is only set up to produce under GLP (Good Laboratory Practice) conditions, which are less stringently regulated than GMP. Lovrecz says that although theoretically his facility could do GMP work, it would limit them to working on one project at a time.

Prof Peter Gray, who runs the Bioengineering Centre at UNSW and is the CEO of the centre's commercial spin-off Acyte, says that his facilities are also limited to pre-clinical bioprocessing. While Acyte concentrates on developing specialised mammalian expression systems for producing pharmaceutical proteins, the Bioengineering Centre covers more of the bioprocessing and manufacturing aspects, with a maximum bioreactor capacity of 140 litres.

A unique approach to bioprocessing is taken by the facility at CSIRO's Food Science Australia, which is part of the CRC for Bioproducts. This facility, which has locations at both North Ryde and Werribee, under the leadership of Dr Philip Franks, works with plant cell cultures for the production of bioactive molecules as well as food products. "Plant material is a source of secondary metabolites, often used as a defence mechanism. As such, these compounds have bioactivity and are often bioactive in humans," explains Franks. "Plant cell culture is good for producing complicated structures which might need several enzymatic steps to synthesise."

Franks says that 25-50 per cent of pharmaceutical products still come from plants. Examples of bioactive plant compounds include aspirin and cancer therapeutic taxol, which is produced in plant cell cultures. "We're doing some work with a company who have an anti-cancer drug to manufacture in plants," he says.

The plant cell culture facility is set up to produce both suspension cultures of de-differentiated plant cells, usually derived from leaf cells which are encouraged to multiply into an undifferentiated state, as well as special root cultures and hairy root cultures which are developed by infecting root cultures with agrobacterium. The facility also has the ability to scale up plant cell cultures from petri dishes to its 10,000 litre fermenter. "The 10,000 litre fermenter is a very large pilot scale. It's probably big enough to kick off a small business," says Franks, noting that 30,000 litres is considered commercial scale.

In addition, the group has specialised equipment to cope with downstream processing, including extraction processes. It is also capable of the more traditional bacterial, fungal and yeast fermentations. While the facility is not yet GMP-compliant, it does meet standards for manufacturing food products, one of the other industries it works with.

"We wouldn't go down that route unless there was a market need for it. We tend to stay on the food side," explains Franks. According to Franks, the group tends to partner with outside companies and institutes, rather than working on internal CSIRO projects. Unlike some of the other facilities, they work with a lot of Australian partners, although through the CRC they have collaborations with groups in China and Japan. "Our primary focus is Australia and Australian industry," he says.

Other facilities have a range of customers and collaborators. CSIRO Molecular Sciences has had customers including Orica, the Ludwig Institute for Cancer Research and Metabolic Pharmaceuticals, as well as one large multinational pharmaceutical company. "We quote quite low by international standards and this should provide us with an advantage, especially with the low Australian dollar," says Zachariou. He notes that the perceived distance of Australia from the Northern Hemisphere biotech hubs is one drawback. "But when you think about what they have to do, it is not a big problem. They only have to send over seed and protocols if any."

Progen has actively sought international contracts, with a stall at this year's Bio 2002 conference in Toronto. "We have contracts in place with two US biotechs and further ones are a possibility," Tillack says, noting that their business is increasingly overseas-based. One of the company's bigger projects is a three-year program funded by the US Malaria Initiative. In collaboration with the Queensland Institute of Medical Research and Monash University, the company is doing all of the process development required to get them up to clinical trials. "We're continually looking for new partnerships -- it's good to be able to draw on other resources," says Tillack.

But if a large part of the customer base for Australia's bioprocessing facilities is not from Australia, what are the reasons for this? Obviously, one is probably the early stage of most Australian biotech start up companies. But there are other reasons, among them perhaps a lack of information on the facilities and services available, and realistic information on the costs of process development services.

"We get an awful lot of requests, do a lot of project proposals, but we get very few that continue. I haven't put my finger on why that is," says Zachariou. He believes that money is one problem - process development is an expensive process.

Bastiras agrees that money for process development is an issue in Australia. "US companies have the money to get the job done quickly," he says.