RE: INTRESTING READ
By Deborah Komlos
Alfalfa has altered his life and provided an alternative approach to recombinant molecule production, and for Louis-Philippe Vézina, PhD there has been no looking back.
Vézina co-founded agbiotech company Medicago Inc. (Ste-Foy, QC) with François Arcand in 1997, creating a firm that uses genetically engineered alfalfa (Medicago sativa L.) lines to produce recombinant proteins for human, veterinary and industrial applications. The privately held company, funded by France-based Avenir Luzerne SA as well as four Quebec venture capitalists, is currently developing its own in-house proteins and has investors keen to support its work on monoclonal antibodies for use in immunotherapy and on plasma proteins.
Inspiration to enter the biotech field came during the mid-’80s, Vézina says, while he was working at the Soils and Crops Research and Development Centre of Agriculture and Agri-Food Canada (AAFC) (Ste-Foy, QC) as a research scientist, a post he obtained before completing his PhD in plant biochemistry at Laval University (Quebec City, QC).
“I got caught up in all the early developments in microbiology for plants,” Vézina says of this period that piqued his interest in biotechnology.
In the next decade, various pressures emerged, he says, such as re-opening of free trade discussions between Europe and North America. Threats subsequently arose that farming subsidies could be cut, especially in Canada. This concern transferred to the milk and beef industries, he says, which rely on forage crops — the area that Vézina was working in.
“We were encouraged to find something to do about this and especially for the plant guys that we were, they just said, try to do something with these plants because they might become useless in a couple of years,” Vézina says, referring to direction the group received from AAFC.
Vézina had been involved in a genetic engineering part of a large alfalfa project at AAFC, and the researchers decided that either they had to find another purpose for the plant, or transform it. They went with the latter, developed the basis for alfalfa transformation technology and tried to sell it to industry. But the task was tough.
“The industry was not ready at all for this,” he says. “It was a very slow emergence of the use of plants to make pharmaceuticals in 1993 or 1994. The first companies were created approximately at that time, but that was a very, very modest effort.”
Making a Change
The stagnant status quo in 1995/96 was Vézina’s catalyst for change.
“I just decided that the industry was not interested,” Vézina says. “I had spent six, seven years developing this and I had spent most of my life working with alfalfa so I thought that something could be done about this. I was at the stage where I just wanted to make a change in my life generally. And I decided to just quit and try to negotiate with Agriculture Canada a licence to use this technology and to bring this to being useful and fruitful economically.”
Vézina says the origins of Medicago, where he is CSO, may not be so different from that of other small startups — negotiating a startup fund with venture capitalists to work out basics such as the intellectual property context, and writing the first business plan. About five or six months after attracting the first round of nearly $1 million in financing, the firm was contacted by Avenir Luzerne SA, an investment company specializing in large-scale production of alfalfa-based animal feed.
“They were looking for a future to their technology and although the GMO (genetically modified organism) context was pretty bad in France, they were interested in what we claimed we could do with the plant,” Vézina says. Avenir Luzerne invested in the company, and the strategic alliance propelled Medicago beyond its competitors, he says, which had comparable molecular biology skills, but lacked the mass-production advantage.
“Getting our hands on industrial-scale technologies from people that had actually grown close to one million hectares of alfalfa and chopping this three, four times a year, crushing the leaves, and performing fractionation in a very, very refined way, this was a very, very big breakthrough for our small industry, not only for Medicago,” Vézina says.
“In one day we became a technology that could really talk about a fully integrated system, which started with the genetic engineering
. . . So, that pushed us quite high in a couple of weeks,” he says.
In 1999, Medicago opened its R&D laboratories and greenhouses. A year later, it formed a Strategic Development Team to begin selling the technology. In 2001, the company implemented its Prototype Development Unit to service the first commercial agreements, and continued doing so in its Dedicated Production Units last year.
Vézina says the firm, which has 45 members including 30 scientists, is trying to close a second round of financing with its investors, and if all goes well, should reach the end of preclinical trials and initiation of Phase I trials at the end of 2005.
Today, fewer than 15 companies worldwide have developed technologies to transform crops such as corn, alfalfa and rice into potentially large-scale protein factories, Vézina says, adding that most do not have the intellectual property and licences required for commercial production.
“We can consider our technology to be a bit of a second wave because it’s very different from what they do,” he says. By “they” he means firms such as Meristem® Therapeutics SA (Clermont-Ferrand, France), ProdiGene Inc. (College Station, TX) and SemBioSys Genetics Inc. (Calgary, AB), which were among the early companies that started circulating and talking to people a couple of years before Medicago hit the scene. “Most of the second-wave companies, in which we are, have developed approaches for the industry which are a lot more comprehensive and easy to use and user-friendly,” he says.
Critical Details
Medicago has created a five-step process to produce recombinant molecules. Initially, a cDNA encoding a protein of interest is inserted into an expression cassette. The firm has developed proprietary cassette constructs, including promoter families, comprising DNA fragments originated from alfalfa, to regulate expression of the inserted gene.
Step two involves alfalfa transformation technology using two possible approaches: Agrobacterium-mediated transformation and protoplast transformation. “It’s a very, very large improvement of the methods that were developed in the 1990s for other species,” he says of protoplast transformation. “We adapted it to alfalfa and one of the key aspects of this is we can actually perform transformation with, I would say, a good yield of transformation, but without using any selection pressure — that means we don’t use any antibiotic marker or anything like that.”
Plant biomass production follows via greenhouse propagation. Next, the protein product is recovered and purified from the leaves, a step that Medicago is actively researching to be able to produce recombinant proteins for a range of product specifications, from unpurified commodities to injectable drugs. Another research focus is meeting homogeneity requirements from national food and drug administrations. In the final step, the firm aims to process residues into byproducts when economically viable.
Alfalfa as a bio-reactor has several benefits, Vézina says. For one, the plant is perennial and is grown easily via stem cuttings, therefore not needing seed production. This also permits selection of individual clones, which permits faster and easier population scale-up to provide the foliage used for the protein expression.
“The dry matter of a corn seed has approximately six per cent protein and the dry matter of an alfalfa leaf has around 22 per cent of protein,” Vézina says. “So if you succeed in producing one per cent of the total protein in corn or in alfalfa you will end up with three times more in alfalfa.”
In addition, he says, certain regions such as southern Ontario can have harvests four times a year, which brings the total harvest of dry matter to around 10, 12, or in some places, 14 tons per hectare. “This is a lot higher than what you can expect from corn or canola or any of the seed plants,” Vézina says.
A challenge faced by Medicago that the seed-firms lack, however, is handling material that is much better fresh than dry, Vézina points out. Although the leaves can be dried, pulverized and still have proteins extracted even after six months of storage, he says this may become a costly procedure at a large scale.
Coming Up
Aside from existing technical concerns, Vézina identifies two regulatory-related hurdles facing Medicago. The first is to gain approval by the U.S. Food and Drug Administration for demonstrating reproducibility in its products and that the preparations lack toxic or immunogenic compounds. There is a common fear in relation to these recombinant products, he says, that plant compounds may produce toxicity or allergenic reactions. The second regulatory hurdle is also the object of fear: growing GMOs in the field.
“If we want to take the real advantage of plants we will have to be able to do these things in large scale, and greenhouses are not really large scale,” Vézina says. But the development of appropriate confinement guidelines and monitoring procedures will hopefully help alleviate this problem, he adds.
A characteristic that has been essential for Medicago’s success to date, and is one that is needed for future progress, is the right outlook. “I wouldn’t be here where I am if I wasn’t an optimist,” Vézina says. “I still believe in this technology.
“I don’t think all these guidelines and confinement and all this will kill the industry,” he says. “It will just reassure everybody and ourselves as well because once the public is reassured, then we will stop fighting about things which are more of the imagination than reality and do the real thing, and if it’s confinement, then let’s do it.”
Although Vézina is no longer an academic scientist or faculty member — before Medicago he was also an associate professor at Laval University — he says he makes sure to maintain a strong scientific presence in the firm.
“I still like doing science and I’m still scientific director here and I could pass on a lot of responsibilities but still, I didn’t do this,” he says. “I still go to the lab most of the days I’m here and I’m still instrumental in doing good science and pushing the technology further.”
Vézina thrives on the competitive aspect of biotech, something he says is not felt in academia, not even from the typical expectation and requirement to churn out publications in academic research environments.
“When you sit in a meeting and you see your competition saying that they’re producing something which you know is 10 times better than what you do and that you have six months to find something else, this is real competition; this is what I like,” he says.
Also enjoyable, Vézina says, is working with people, and his colleagues share his enthusiasm for the work. “They’re all eager to perform . . . We’ve been through very tough times and they’re still there. They’re still very, very motivated,” he says.
When asked if he ever regrets his decision years ago to change his life and career path by pursuing alfalfa into the biotech world, the resolve in Vézina’s reply could probably not be greater:
“Oh, no, no, no, not in a million years.”