Feature: Manufacturing monoclonal antibodies

The journey to market for biological products can be a complex and frustrating process. A key challenge in this process is producing or obtaining the relevant compounds in quantities large enough to support research studies, pre-clinical studies and/or early-stage clinical trials.

Projects often slow or even stop along the journey as they struggle at this step. A few years ago, the federal government committed $35 million to address just this problem as a part of the National Collaborative Research Infrastructure Strategy (NCRIS).

Subsequently, NCRIS put the call out for proposals that would enhance Australia’s capacity for large-scale biotechnology production. The funding would cover three distinct areas – recombinant proteins for use as potential therapeutics, human cells for transplantation or clinical activity involving human subjects, and biofuels – the ultimate aim being to establish a national infrastructure that would help Australian biotechnology reach globally competitive levels.

One team who successfully secured a piece of this NCRIS action was that led by Dr Chris Marquis, who is director of the recently established Recombinant Products Facility (RPF) at the University of New South Wales.

According to Marquis, groups were set up all over the place to bid for this new pot of money, broken up into 12 Capabilities, covering everything from biomolecular platforms, ecology, radio astronomy and biotechnology.

“In the biotechnology area, one of the capabilities identified as a priority need was a network of national infrastructure so that researchers could access the production and purification of proteins at a price that would be lower than that charged commercially – in other words, affordable to them,” says Marquis.

“The ones that got up were those that basically had a cohesive story to tell and could leverage more funding out of the state government and their own institutions or companies etc. Successful proposals also had to show some involvement and potentially some personal access to facilities and staff, rather than a hands-off contract arrangement.”

And so, in 2009, the RPF at the University of New South Wales was born. The facility grew out of an existing bioprocessing laboratory set up over 15 years ago as a small- to medium-scale animal culture facility by Professor Peter Gray.

The laboratory already provided cell line development, bioprocess development and protein production services to the research community and industry, and so was perfectly situated to step up to the NCRIS challenge.

The facilities at the University of New South Wales were subsequently upgraded using the NCRIS funding, and other nodes of the Capability were established at the Australian Institute of Bioengineering and Nanotechnology (AIBN) at The University of Queensland (now headed by Peter Gray) and at a Victorian-based facility established jointly by Monash University’s ARC Special Research Centre for Green Chemistry and the CSIRO’s Division of Molecular and Health Technologies.

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Down to business

“Our facility at UNSW now has more of a microbial focus than the existing facility used to, although we do still run some cell culture projects and hybridoma work,” says Marquis.

“Under the NCRIS Capability we provide subsidised access to our facilities for public and private sector research, and the funding allowed us to upgrade our capacity in all areas – most importantly it also allowed us to hire the staff to do that and to actually run the facility as prescribed by the NCRIS strategy.”

Since expanding, the RPF now provides a range of services, including cell line and hybridoma development and characterisation, bioprocess development, microbial fermentation up to 25 litres, mammalian cell culture to 10 litres, protein purification and analysis and cell line storage (-80° C and liquid nitrogen).

The other NCRIS nodes fill slightly different niches for the Capability, specialising in protein expression (AIBN) and all manner of downstream processing (AIBN and CSIRO).

A key principle of NCRIS, according to Marquis, is that the funded facilities have to be accessible on the basis of merit at reasonable prices, wherever they are located in Australia.

To this end, researchers needing significant quantities of proteins through the RPF for specific purposes such as clinical trials may also qualify for subsidised access to the licensed contract manufacturing organisations, Hospira Adelaide and RadPharm Scientific in Canberra (a division of Global Medical Solutions).

Marquis reports that the facilities at UNSW see a wide range of users through the doors. A lot are researchers from academic or research institutions such as the Garvan and Heart Research Institutes in Sydney, while other users are from biotech companies.

Under the NCRIS guidelines and pricing/access rules, preference is given to those commercial enterprises with less than 15 employees, and so far the RPF has worked with several such small start-up companies, some from Brisbane and others in Sydney.

Happy customers

In a recent project, the RPF purified large amounts of a native protein that was just entering early-stage trials for a biomedical application – according to Marquis, they purified over 150 grams of the stuff.

“In this sort of example, the key challenges in producing proteins for in-human trials are to bring as close to a good manufacturing practice [GMP] environment to the job as possible, without actually being an accredited GMP facility, and this was a steep learning the first time we had to do it, although it is much easier now.”

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Another project that is just about to be published involved developing and producing a monoclonal antibody to be used in pre-clinical animal trials in the anti-cancer area. “This work looks really interesting, although I can’t name names just yet.”

One of Marquis’ own research interests also forms an ongoing part of the RPF activities. Together with Dr Alice Lee in the School of Chemical Engineering at UNSW, Marquis is interested in purifying food allergens, specifically those derived from peanuts.

Peanut allergy is a common and severe food allergy in Western societies, and efforts to improve allergy testing and develop treatments require the relevant proteins from peanuts – recombinant and native versions – in large quantities.

Marquis is focusing on expressing and purifying recombinant versions of some of the major peanut-derived proteins that have been identified to cause this mega-allergic response in susceptible individuals.

The ultimate aim is to produce monoclonal antibodies in mice to a selection of these peanut antigens and use them in immunoassays for the food industry and clinical use, and potentially to develop treatment strategies.

“We are also looking at ways to make standard recombinant versions of these native proteins so they can be used as diagnostic standards. The plants naturally produce variant forms of such proteins depending on genetic and environmental elements so making a standard for that protein is not as simple as it seems,” he says.”

“Through the cell culture facility, I have also done other collaborative work looking at things like interfacing with nanomaterials for applications in gene/RNA delivery and nanotoxicity-type studies.

This is a bit to the left of the recombinant proteins area, but we use some of the facilities here at the Capability to support such activities through collaborative work.”

This feature appeared in the September/October 2010 issue of Australian Life Scientist. To subscribe to the magazine, go here.